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In this study, we present an explicit crystal plasticity finite element model (CPFEM) to investigate the texture evolution during multipass tube spinning of a TA15 alloy. The texture formation mechanism is further revealed by analyzing grain rotation and slip behaviors. The results indicate that, for the grains whose c-axes are far from the normal direction (ND), the basal slip is dominant under the significant compression deformation state of spinning. Therefore, the c-axis gradually turns toward the ND. However, for the grains whose c-axes are close to the ND, the contribution ratios of other slip systems increase, especially that of prism . Under these conditions, the c-axis deviates slightly from the ND, and grains mainly rotate around the c-axis. Therefore, most of the {0001} poles cluster around the ND and {10-10} poles are distributed in the rolling-hoop plane, exhibiting an obvious basal texture component. These results imply that the CPFEM can effectively predict the formation of basal texture component during spinning. With an increase in the number of spinning pass, the texture distribution becomes slightly inhomogeneous along the ND owing to the uneven distribution of macroscopic strain. In addition, the grain rotation first increases and then decreases with deformation. The smaller grain rotation at the first pass is because a large number of basal preferentially operates in the grain with favorable orientation, resulting in localized inhomogeneous deformation. The decrease at larger deformation is due to the lower Schmidt factor of basal , which limits the grain rotation.

In this study, the texture formation mechanism of a TA15 titanium alloy under different plane strain compression conditions was investigated by analyzing the slipping, dynamic recrystallization (DRX) and phase transformation behaviors. The results indicated that the basal texture component basically appears under all conditions, since the dominant basal slip makes the C-axis of the α grain rotate to the normal direction (ND, i.e., compression direction), but it has a different degree of deflection. With an increase in deformation amount, temperature or strain rate, {0001} poles first approach the ND and then deviate from it. Such deviation is mainly caused by a change in slip behaviors and phase transformation. At a smaller deformation amount and higher strain rate, inhomogeneous deformation easily causes a basal slip preferentially arising from the grain with a soft orientation, resulting in a weak basal texture component. A greater deformation amount can increase the principal strain ratio, thereby promoting other slip systems to be activated, and a lower temperature can increase the critical shear stress of the basal slip, further causing a dispersive orientation under these conditions. At a higher temperature and a lower strain rate, apparent phase transformation will induce the occurrence of lamellar α whose orientation obeys the Burgers orientation of the β phase, thereby disturbing and weakening the deformation texture. As for DRX, continuous-type (CDRX) is most common under most conditions, whereas CDRX grains have a similar orientation to deformed grains, so DRX has little effect on overall texture. Moreover, the microhardness of samples is basically inversely proportional to the grain size, and it can be significantly improved as lamellar α occurs. In addition, deformed samples with a weaker texture present a higher microhardness due to the smaller Schmidt factors of the activated prism slip at ambient loading.

The correlation between magnetic Barkhausen noise (MBN) features and the surface hardness of two types of die steels (Cr12MoV steel and S136 steel in Chinese standards) was investigated in this study. Back-propagation neural network (BP-NN) models were established with MBN magnetic features extracted by different methods as the input nodes to realize the quantitative prediction of surface hardness. The accuracy of the BP-NN model largely depended on the quality of the input features. In the extraction process of magnetic features, simplifying parameter settings and reducing manual intervention could significantly improve the stability of magnetic features. In this study, we proposed a method similar to the magnetic Barkhausen noise hysteresis loop (MBNHL) and extracted features. Compared with traditional MBN feature extraction methods, this method simplifies the steps of parameter setting in the feature extraction process and improves the stability of the features. Finally, a BP-NN model of surface hardness was established and compared with the traditional MBN feature extraction methods. The proposed MBNHL method achieved the advantages of simple parameter setting, less manual intervention, and stability of the extracted parameters at the cost of small accuracy reduction.

Mangrove forests inhabit tropical or subtropical intertidal zones and have remarkable abilities in coastline protection. Kandelia obovata is considered the most cold-tolerant mangrove species and has been widely transplanted to the north subtropical zone of China for ecological restoration. However, the physiological and molecular mechanisms of K. obovata under colder climate was still unclear. Here, we manipulated the typical climate of cold waves in the north subtropical zone with cycles of cold/recovery and analyzed the physiological and transcriptomic responses of seedlings. We found that both physiological traits and gene expression profiles differed between the first and later cold waves, indicating K. obovata seedlings were acclimated by the first cold experience and prepared for latter cold waves. 1,135 cold acclimation-related genes (CARGs) were revealed, related to calcium signaling, cell wall modification, and post-translational modifications of ubiquitination pathways. We identified the roles of CBFs and CBF-independent transcription factors (ZATs and CZF1s) in regulating the expression of CARGs, suggesting both CBF-dependent and CBF- independent pathways functioned in the cold acclimation of K. obovata. Finally, we proposed a molecular mechanism of K. obovata cold acclimation with several key CARGs and transcriptional factors involved. Our experiments reveal strategies of K. obovata coping with cold environments and provide prospects for mangrove rehabilitation and management.

Abstract Most of the biological processes, including cell signaling, cancer invasion, embryogenesis, or neural development, are dependent on and guided by the complex architecture and composition of cellular microenvironments. Mimicking such microenvironments in cell coculture models is crucial for fundamental and applied biology investigations. The ability to combine different cell types grown as both two‐dimensional (2D) monolayers and three‐dimensional (3D) spheroids in specific defined location inside a microculture environments is a key towards in vitro tissue modeling and towards mimicking complex in vivo cellular processes. In this study, we introduce and investigate a method to create in vitro models of 2D cell monolayers cocultured with 3D spheroids in defined preorganization. We demonstrate the possibility of creating such complex cellular microenvironments in a high‐throughput and automated manner by creating arrays of such droplets containing prearranged 2D and 3D cellular microcolonies. Furthermore, we demonstrate an application of this approach to study paracrine propagation of Wnt signaling between 2D and 3D cellular colonies. This method provides a general approach for the miniaturized, high‐throughput, and automated formation of complex coculture cellular microarchitectures that will be useful for mimicking various in vivo complex cellular structures and for studying complex biological processes in vitro.

The allocation of biomass reflects a plant’s resource utilization strategy and is significantly influenced by climatic factors. However, it remains unclear how climate factors affect the aboveground and belowground biomass allocation patterns on macro scales. To address this, a study was conducted using aboveground and belowground biomass data for 486 species across 294 sites in China, investigating the effects of climate change on biomass allocation patterns. The results show that the proportion of belowground biomass in the total biomass (BGBP) or root-to-shoot ratio (R/S) in the northwest region of China is significantly higher than that in the southeast region. Significant differences (p < 0.05) were found in BGBP or R/S among different types of plants (trees, shrubs, and herbs plants), with values for herb plants being significantly higher than shrubs and tree species. On macro scales, precipitation and soil nutrient factors (i.e., soil nitrogen and phosphorus content) are positively correlated with BGBP or R/S, while temperature and functional traits are negatively correlated. Climate factors contribute more to driving plant biomass allocation strategies than soil and functional trait factors. Climate factors determine BGBP by changing other functional traits of plants. However, climate factors influence R/S mainly by affecting the availability of soil nutrients. The results quantify the productivity and carbon sequestration capacity of terrestrial ecosystems and provide important theoretical guidance for the management of forests, shrubs, and herbaceous plants.

Changes in net primary productivity (NPP) to global change have been studied, yet the relative impacts of global change on grassland productivity at large scales remain poorly understood. Using 182 grassland samples established in 17 alpine meadows (AM) and 21 desert steppes (DS) in China, we show that NPP of AM was significantly higher than that of DS. NPP increased significantly with increasing leaf nitrogen content (LN) and leaf phosphorus content (LP) but decreased significantly with increasing leaf dry matter content (LDMC). Among all abiotic factors, soil nutrient factor was the dominant factor affecting the variation of NPP of AM, while the NPP of DS was mainly influenced by the changing of precipitation. All abiotic factors accounted for 62.4% of the spatial variation in the NPP of AM, which was higher than the ability to explain the spatial variation in the NPP of DS (43.5%). Leaf traits together with soil nutrients and climatic factors determined the changes of the grassland productivity, but the relative contributions varied somewhat among different grassland types. We quantified the effects of biotic and abiotic factors on grassland NPP, and provided theoretical guidance for predicting the impacts of global change on the NPP of grasslands.

Plant functional traits are a representation of plant resource utilization strategies. Plants with higher specific leaf area (SLA) and lower leaf dry matter content (LDMC) exhibit faster investment-return resource utilization strategies. However, the distribution patterns and driving factors of plant resource utilization strategies at the macroscale are rarely studied. We investigated the relative importance of climatic and soil factors in shaping plant resource utilization strategies at different life forms in forests using data collected from 926 plots across 163 forests in China. SLA and LDMC of plants at different life forms (i.e., trees, shrubs, and herbs) differ significantly. Resource utilization strategies show significant geographical differences, with vegetation in the western arid regions adopting a slower investment-return survival strategy and vegetation in warmer and wetter areas adopting a faster investment-return survival strategy. SLA decreases significantly with increased temperature and reduced rainfall, and vegetation growing in these conditions exhibits conservative resource utilization. Mean annual precipitation (MAP) is a key climatic factor that controls the resource utilization strategies of plants at the macroscale. Plants use resources more conservatively as soil pH increases. The influence of climate and soil factors is coupled to determine the resource utilization strategies of plants occupying different life forms at the macroscale, but the relative contribution of each varies across life forms. Our findings provide a theoretical framework for understanding the potential impact of increasing global temperatures on plant resource utilization.

Reception of Wnt signals by cells is predominantly mediated by Frizzled receptors in conjunction with a co-receptor, the latter being LRP6 or LRP5 for the Wnt/β-catenin signalling pathway. It is important that cells maintain precise control of receptor activation events in order to properly regulate Wnt/β-catenin signalling as aberrant signalling can result in disease in humans. Phosphorylation of the intracellular domain (ICD) of LRP6 is well known to regulate Wntβ-catenin signalling; however, less is known for regulatory post-translational modification events within the extracellular domain (ECD). Using a cell culture-based expression screen for functional regulators of LRP6, we identified a glycosyltransferase, B3GnT2-like, from a teleost fish (medaka) cDNA library, that modifies LRP6 and regulates Wnt/β-catenin signalling. We provide both gain-of-function and loss-of-function evidence that the single human homolog, B3GnT2, promotes extension of polylactosamine chains at multiple N-glycans on LRP6, thereby enhancing trafficking of LRP6 to the plasma membrane and promoting Wnt/β-catenin signalling. Our findings further highlight the importance of LRP6 as a regulatory hub in Wnt signalling and provide one of the few examples of how a specific glycosyltransferase appears to selectively target a signalling pathway component to alter cellular signalling events.

The combination of multifunctional micromagnetic testing and neural network-based prediction models is a promising way of nondestructive and quantitative measurement of steel surface hardness. Current studies mainly focused on improving the prediction accuracy of intelligent models, but the unavoidable and random uncertainties related to instruments were seldom explored. The robustness of the prediction model considering the repeatability of instruments was seldom discussed. In this work, a self-developed multifunctional micromagnetic instrument was employed to perform the repeatability test with Cr12MoV steel. The repeatability of the instrument in measuring multiple magnetic features under both static and dynamic conditions was evaluated. The magnetic features for establishing the prediction model were selected based on the consideration of both the repeatability of the instrument and the ability of magnetic features in surface hardness evaluation. To improve the robustness of the model in surface hardness prediction, a modelling strategy considering the repeatability of the instrument was proposed. Through removing partial magnetic features with higher mean impact values from input nodes, robust evaluation of surface hardness in Cr12MoV steel was realized with the multifunctional micromagnetic instrument.

Magnetic Barkhausen noise (MBN), sensitive to the microstructure of materials, can be applied in the surface decarburization depth detection of ferromagnetic specimens. However, the effects of core microstructures on the determination results of decarburization depth have not been explored. In this study, MBN was employed to evaluate the magnetic properties of the decarburized 60Si2Mn spring steels with martensitic and pearlitic core microstructures. Spring steel samples were austenitized at different times to generate different decarburization depths. Seven magnetic features were extracted from the MBN butterfly profiles. We used the variation coefficient, linear correlation coefficient, and normalized sensitivity to discuss the influence of the core microstructures on these seven features. The different core microstructures led to a large difference in the ability of MBN features to characterize the decarburization layer depth. However, three features of MBN butterfly profiles demonstrated an approximately linear dependency (linear correlation coefficient > 94%) on surface decarburization depth and monotonically increased with the increase in depth in both core microstructures of spring steels.

Bacillus subtilis J-15 is a plant growth-promoting rhizobacteria isolated from the soil rhizosphere of cotton and is resistant to cotton verticillium wilt. This study evaluated the effects of metabolites of J-15 (J-15-Ms), including mycosubtilin, on plant growth using Arabidopsis and cotton plants. The results showed that J-15-Ms promoted Arabidopsis seeding growth at lower concentrations of 0.2 μg/mL but inhibited the growth at higher concentrations, such as 20 μg/mL. Similar results were obtained in cotton. Thus, J-15-Ms-treated plants showed low-concentration-induced growth promotion and high-concentration-induced growth inhibition. The J-15-Ms components were analyzed by liquid chromatography–mass spectrometry. Correlation analysis using the J-15 genomic databases suggested that J-15 may synthesize indoleacetic acid via the indole-3-pymvate pathway and indole-3-acetamide pathway. Treatment with mycosubtilin, a purified peptide from J-15-Ms, showed that the peptide promoted Arabidopsis growth at a low concentration (0.1 μg/mL) and inhibited plant growth at high concentrations (higher than 1 μg/mL), which also significantly increased plant lateral root number. Transcriptomic analysis showed that mycosubtilin might promote lateral root development and inhibit plant primary root growth by regulating the expression of the plant hormone signaling pathway. This study reveals the mechanism of Bacillus subtilis J-15 in affecting plant growth.

Leaf nutrient content and its stoichiometric relationships (N/P ratio) are essential for photosynthesis and plant growth and development. Previous studies on leaf nutrient-related functional traits have mainly focused on the species level and regional scale, but fewer studies have investigated the distribution patterns of the leaf N and P contents (LN, LP) and N/P ratios (N/P) in communities and their controlling factors at a large scale; therefore, we used LN, LP, and N/P data at 69 sites from 818 forests in China. The results showed significant differences (p < 0.05) in the LN, LP, and N/P at different life forms (tree, shrub, and herb). Neither LN, LP, nor N/P ratios showed significant patterns of latitudinal variation. With the increase in temperature and rainfall, the LN, LP, and leaf nutrient contents increased significantly (p < 0.001). Across life forms, LN at different life forms varied significantly and was positively correlated with soil P content (p < 0.001). The explanatory degree of climatic factors in shaping the spatial variation patterns of LN and N/P was higher than that of the soil nutrient factors, and the spatial variation patterns of the leaf nutrient traits of different life forms were shaped by the synergistic effects of climatic factors and soil nutrient factors.

Glucose metabolism regulates cell growth and affects astaxanthin accumulation in the green algae Chromochloris zofingiensis. Hub gene functioning in this bioactive compound has been illustrated at the genome, transcriptome and metabolome level, but is rather limited from a proteome aspect. Microalgal cell produce an enhanced biomass (8-fold higher) but decreased lipid and astaxanthin content (~20% less) in the glucose condition compared to the control. Here, we investigate the proteomic response of C. zofingiensis grown with and without glucose using an LC-MS/MS-based Tandem Mass Tag (TMT) approach. The proteomic analysis demonstrated that glucose supplementation triggers the upregulation of 105 proteins and downregulation of 151 proteins. Thus, the carbon and energy flux might flow to cell growth, which increased the associated protein abundance, including DNA polymerase, translation initiation factor, 26S proteasome regulatory subunits, and the marker enzyme of the TCA cycle ribosomal protein. Moreover, the glucose supplement triggered the downregulation of proteins mainly involved in photosynthesis, chloroplasts, valine, leucine and isoleucine biosynthesis, 2-oxocarboxylic acid metabolism, and pantothenate and CoA biosynthesis pathways. This proteomic analysis is likely to provide new insights into algal growth and lipid or astaxanthin accumulation upon glucose supplementation, providing a foundation for further development of C. zofingiensis as oleaginous microalga for bioengineering applications.

Development and homeostasis of multicellular organisms is largely controlled by complex cell-cell signaling networks that rely on specific binding of secreted ligands to cell surface receptors. The Wnt signaling network, as an example, involves multiple ligands and receptors to elicit specific cellular responses. To understand the mechanisms of such a network, ligand-receptor interactions should be characterized quantitatively, ideally in live cells or tissues. Such measurements are possible using fluorescence microscopy yet challenging due to sample movement, low signal-to-background ratio and photobleaching. Here, we present a robust approach based on fluorescence correlation spectroscopy with ultra-high speed axial line scanning, yielding precise equilibrium dissociation coefficients of interactions in the Wnt signaling pathway. Using CRISPR/Cas9 editing to endogenously tag receptors with fluorescent proteins, we demonstrate that the method delivers precise results even with low, near-native amounts of receptors.

Microalgal cells serve as solar-powered factories that produce pharmaceuticals, recombinant proteins (vaccines and drugs), and valuable natural byproducts that possess medicinal properties. The main advantages of microalgae as cell factories can be summarized as follows: they are fueled by photosynthesis, are carbon dioxide-neutral, have rapid growth rates, are robust, have low-cost cultivation, are easily scalable, pose no risk of human pathogenic contamination, and their valuable natural byproducts can be further processed. Despite their potential, there are many technical hurdles that need to be overcome before the commercial production of microalgal pharmaceuticals, and extensive studies regarding their impact on human health must still be conducted and the results evaluated. Clearly, much work remains to be done before microalgae can be used in the large-scale commercial production of pharmaceuticals. This review focuses on recent advancements in microalgal biotechnology and its future perspectives.

Wild soybean (Glycine soja Sieb.et Zucc; WS) has been used as a traditional food in China for many years and contains significantly higher levels of isoflavones than cultivated soybean (Glycine max; CS), but the secondary metabolites, including flavonoids and the phenolic composition differences between them, remain unclear. The results showed that WS possessed significantly higher total phenolic and flavonoid content and exhibited better antioxidant and α-glucosidase inhibition activities as well as excellent protective effects against H2O2-induced oxidative injury in a human endothelial cell line. Through metabolomic analysis, 642 metabolites were identified, and 238 showed differential expression, with 151 upregulated and 87 downregulated. A total of 79 flavonoid compounds were identified, 42 of which were upregulated in WS. 2′-Hydroxygenistein, garbanzol, protocatechuic aldehyde, ligustilide, and resveratrol were the most discriminated compounds in WS. The metabolic pathway analysis of differential metabolites related to the biosynthesis of flavonoids and phenolic acids were the biosynthesis of phenylpropanoids, flavonoids, isoflavonoids, flavones, and flavonols. This study substantially elucidated differences in the content of flavonoids and biological activities between WS and CS, which is useful information for the effective utilization of these two black soybean species in food processing.

The device-to-device (D2D) communication has been regarded as an effective technique for complementing and enhancing the conventional cellular systems owing to its capability of substantially improving both the spectral and power efficiencies of wireless networks. However, the severe interference imposed on the conventional cellular users (CUs) by the geographically close-by D2D pairs may cause a significant performance erosion in the D2D-aided underlaying cellular networks (CNs). In this paper, performance analysis for the D2D-aided underlaying CNs in terms of throughput is provided. We first derive the closed-form expressions of the coverage probability for both the conventional cellular links and the D2D links, followed by giving out the approximated expressions of the ergodic data rate for both an individual cellular/D2D link and the whole underlaying network. Furthermore, the key parameters (e.g., the density of D2D users (DUs) or CUs, and the average geographical distance between a pair of D2D peers) significantly impacting the channel capacity are adaptively adjusted for maximizing the sum data rate of the proposed underlaying networks. In addition, both theoretical analysis and simulation results reveal the attainability of the maximal throughput by optimizing the critical parameters, such as the density of DUs, provided that the scale factor between the DUs and sum users (i.e., comprising both conventional CUs and DUs) can be effectively balanced subject to the constraints specified in the proposed scheme.

Tube spinning is an effective plastic-forming technology for forming light-weight, high-precision and high-reliability components in high-tech fields, such as aviation and aerospace. However, cracks commonly occur in tube spinning due to the complexity of stress state, which severely restricts the improvement of the forming quality and forming limit of components. In this study, a finite element (FE) model coupled with Gurson-Tvergaard-Needleman (GTN) damage model for forward tube spinning of 3A21-O aluminum alloy is established and its applicability is evaluated by experiment. Meanwhile, the GTN damage model is employed to study the damage evolution for forward tube spinning of 3A21-O aluminum alloy. The results show that the FE model is appropriate for predicting the macroscopic crack appearing in uplift area for forward tube spinning, while the damage evolution in deformation area could not be predicted well due to the negative stress triaxiality and the neglect of shear deformation. Accumulation of damage in forward tube spinning occurs mainly in the uplift area. Void volume fraction (VVF) in the outer surface of the tube is higher than that in the inner surface. In addition, it is prone to cracking in the outer surface of tube in the material uplift area.

Due to the restrictions of traditional teaching model, serious problems emerge in Comprehensive English teaching, which hamper the practical competence of English major students. Based on the essentials of John Dewey's pragmatic ideas, this study explores and proposes the 3-dimentional teaching model in Comprehensive English and aims to promote English basic skills, innovative thinking intelligence and professional practical competence of the English major talents. [ABSTRACT FROM AUTHOR]

Tools such as agricultural grass cutters, stubble blades, machetes, and grain crushing knives can become passivated gradually during use without self-sharpening. To resolve this problem, a composite coating of a Ni-based alloy and tungsten carbide (WC) was brazed onto the unilateral surface of 45 steel tools. To ensure that the unilateral coating tool can self-sharpen, different heat treatments were tested that can change the microstructure of the substrate. This makes it possible to regulate the wear resistance on both sides of the tool. Previous research shows that multiple phases can coexist at the interface between WC and Ni-based brazing alloys. These can decrease the initial melting temperature and the finishing melting temperature of the coating by 30.7°C and 49°C, respectively. A quantitative wear test of the tool indicates that the material-removal rate of the coating and the substrate is inversely proportional to its hardness. As a result, the optimal coating thickness can be estimated using the material-removal rate, i.e., the wear. A plowing simulation in gravel shows that after long-time usage (for example, 100 h), the tool, which was treated with quenching/tempering, is very good at self-sharpening. Moreover, the shortening in the direction of the symmetry line of the tool is small. The aim of the study is to improve both self-sharpening and service life of the tools. [ABSTRACT FROM AUTHOR]

Resource allocation in device-to-device (D2D) aided cellular systems, in which the proximity users are allowed to communicate directly with each other without relying on the intervention of base stations (BSs), is investigated in this paper. A new uplink resource allocation policy is proposed by exploiting the relationship between D2D-access probability and channel gain among variant devices, such as cellular user equipments (CUEs), D2D user equipments (DUEs) and BSs, etc., under the constraints of their minimum signal to interference-plus-noise ratio (SINR) requirements. Furthermore, the proposed resource-allocation problem can be formulated as the cost function of "maximizing the number of simultaneously activated D2D pairs subject to the SINR constraints at both CUEs and DUEs". Numerical results relying on system-level simulations show that the proposed scheme is capable of substantially improving both the D2D-access probability and the network throughput without sacrificing the performance of conventional CUEs. [ABSTRACT FROM AUTHOR]

This paper reveals the definition, connotation and significance of critical thinking. It further improves and explores approaches to develop students' critical thinking capability through Comprehensive English teaching and verifies the validity of those approaches through Comprehensive English class implementation and students' class presentation. However, it also suggests that developing critical thinking of instructors shall be prioritized before cultivating that of students', which is worth reflecting in the long run. [ABSTRACT FROM AUTHOR]

This paper analyzes problems in Comprehensive English teaching in light of John Dewey's Pragmatic education philosophy and proposes suggestions to reform Comprehensive English teaching from the perspective of Pragmatic thinking intelligence cultivation, the student-centered approach and proactive assignment outlook. [ABSTRACT FROM AUTHOR]

The Wingless/Int1 (Wnt) signaling system plays multiple, essential roles in embryonic development, tissue homeostasis, and human diseases. Although many of the underlying signaling mechanisms are becoming clearer, the binding mode, kinetics, and selectivity of 19 mammalianWNTs to their receptors of the class Frizzled (FZD1-10) remain obscure. Attempts to investigate Wnt-FZD interactions are hampered by the difficulties in working with Wnt proteins and their recalcitrance to epitope tagging. Here, we used a fluorescently tagged version of mouse Wnt-3a for studying Wnt-FZD interactions. We observed that the enhanced GFP (eGFP)-tagged Wnt-3a maintains properties akin to wild-type (WT) Wnt-3a in several biologically relevant contexts. The eGFP-tagged Wnt-3a was secreted in an evenness interrupted (EVI)/Wntless-dependent manner, activated Wnt/b-catenin signaling in 2D and 3D cell culture experiments, promoted axis duplication in Xenopus embryos, stimulated lowdensity lipoprotein receptor-related protein 6 (LRP6) phosphorylation in cells, and associated with exosomes. Further, we used conditioned medium containing eGFP-Wnt-3a to visualize its binding to FZD and to quantify Wnt-FZD interactions in real time in live cells, utilizing a recently established NanoBRETbased ligand binding assay. In summary, the development of a biologically active, fluorescent Wnt-3a reported here opens up the technical possibilities to unravel the intricate biology of Wnt signaling and Wnt-receptor selectivity. [ABSTRACT FROM AUTHOR]