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Organophosphorus flame retardants (OPFRs) are important alternatives to brominated flame retardants (BFRs), but information on their contamination of the environment in China is rare. We examined the occurrence of 12 OPFRs in indoor dust in four microenvironments of southern China, including a rural electronic waste (e-waste) recycling area, a rural non-e-waste area, urban homes, and urban college dormitory rooms. The OPFR concentrations (with a median of 25.0 μg g−1) were highest in the e-waste area, and the concentrations in other three areas were lower and comparable (7.48–11.0 μg g−1). The levels of OPFRs in the present study were generally relatively lower than the levels of OPFRs found in Europe, Canada, and Japan because BFRs are still widely used as the major FRs in China. The composition profile of OPFRs in the e-waste area was dominated by tricresyl phosphate (TCP) (accounting for 40.7%, on average), while tris(2-chloroethyl) phosphate (TCEP) was the most abundant OPFR (64.4%) in the urban areas (homes and college dormitories). These two distribution patterns represent two OPFR sources (i.e., emissions from past e-waste and from current household products and building materials). The difference in the OPFR profiles in the rural area relative to the OPFR profiles in the urban and e-waste areas suggests that the occurrence of OPFRs is due mainly to emissions from characteristic household products in rural homes. Although human exposures to all the OPFRs were under the reference doses, the health risk for residents in the e-waste area is a concern, considering the poor sanitary conditions in this area and exposure from other sources.

Protein arginine methyltransferase 1 (PRMT1), a type-I arginine methyltransferase, has been implicated in diverse cellular events. We have focused on the role of PRMT1 in gliomagenesis. In this study, we showed that PRMT1 expression was up-regulated in glioma tissues and cell lines compared with normal brain tissues. The knock-down of PRMT1 resulted in an arrest in the G1-S phase of the cell cycle, proliferation inhibition and apoptosis induction in four glioma cell lines (T98G, U87MG, U251, and A172). Moreover, an in vivo study confirmed that the tumor growth in nude mouse xenografts was significantly decreased in the RNAi-PRMT1 group. Additionally, we found that the level of the asymmetric dimethylated modification of H4R3, a substrate of PRMT1, was higher in glioma cells than in normal brain tissues and decreased after PRMT1 knock-down. Our data suggest a potential role for PRMT1 as a novel biomarker of and therapeutic target in gliomas. [BMB Reports 2012; 45(8): 470-475]

MicroRNAs (miRNAs) are small noncoding regulatory RNAs whose aberrant expression may be observed in many malignancies. However, few data are yet available on human primary medulloblastomas. This work aimed to identify that whether miRNAs would be aberrantly expressed in tumor tissues compared with non-tumorous cerebellum tissues from same patients, and to explore a possible role during carcinogenesis.A high throughput microRNA microarray was performed in human primary medulloblastoma specimens to investigate differentially expressed miRNAs, and some miRNAs were validated using real-time quantitative RT-PCR method. In addition, the predicted target genes for the most significantly down- or up-regulated miRNAs were analyzed by using a newly modified ensemble algorithm.Nine miRNA species were differentially expressed in medulloblastoma specimens versus normal non-tumorous cerebellum tissues. Of these, 4 were over expressed and 5 were under expressed. The changes ranged from 0.02-fold to 6.61-fold. These findings were confirmed using real-time quantitative RT-PCR for most significant deregulated miRNAs (miR-17, miR-100, miR-106b, and miR-218) which are novel and have not been previously published. Interestingly, most of the predicted target genes for these miRNAs were involved in medulloblastoma carcinogenesis.MiRNAs are differentially expressed between human medulloblastoma and non-tumorous cerebellum tissue. MiRNAs may play a role in the tumorigenesis of medulloblastoma and maybe serve as potential targets for novel therapeutic strategies in future.

To explore the influences of the restoration of neural adhesion molecule NECL1 on the morphology, migration, and invasion of NECL1-deficient glioma cell lines.Scratch and Transwell assays were used to observe the cell migration and invasion, the activities of extracellular metalloproteinases were measured, and the cell morphology was observed. Astrocytes marker glial fibrillary acidic protein was detected by Western blot after the restoration of NECL1 in glioma U251 cell line.In NECL1-deficient U251 glioma cell lines, migration and invasion were inhibited. The U251 cells was differentiated potentially to astrocytes, and glial fibrillary acidic protein was up-regulated after the restoration of the NECL1 expression.As a potential tumor repressor, the neural adhesion molecule NECL1 can inhibit the migration and invasion of glioma cell and induces its differentiation.

Phytochelatins (PCs) play an important role in detoxification of heavy metals in plants. PCs are synthesized from glutathione by phytochelatin synthase (PCS), a dipeptidyltransferase. Sesbania rostrata is a tropical legume plant that can tolerate high concentrations of Cd and Zn. In this study, the S. rostrata PCS gene (SrPCS) and cDNAs were isolated and characterized. Southern blot and sequence analysis revealed that a single copy of the SrPCS gene occurs in the S. rostrata genome, and produces four different SrPCS mRNAs and proteins, SrPCS1-SrPCS4, by alternative splicing of the SrPCS pre-mRNA. The SrPCS1 and SrPCS3 proteins conferred Cd tolerance when expressed in yeast cells, whereas the SrPCS2 and SrPCS4 proteins, which lack the catalytic triad and the N-terminal domains, did not. These results suggested that SrPCS1 and SrPCS3 have potential applications in genetic engineering of plants for enhancing heavy metal tolerance and phytoremediation of contaminated soils.

To study the expression pattern of Polycomb gene Nspc1 at the early developmental stage in zebrafish.In situ hybridization probe for Nspc1 was designed according to the GenBank information. Collecting zebrafish embryos at different stages including one cell stage, two-cell stage, bud stage, and somites stage, we hybridized them with the prepared probe. Then the hybridization signals at different intervals were observed and photographed at the right time.Nspc1 was expressed globally at the early stage. Its expression specificity began at the somites stage, mainly in the nervous system of the head.Nspc1 may play essential roles in the early stage development of zebrafish, especially in the nervous system.

To study the regulation role of tumor suppressor NECL1 on the proliferation of glioma cell line.We detected the expression level of NECL1 in human normal brain tissue and glioma cell lines using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot. T98G cell line in which NECL1 was silent and whose transfection efficiency was relatively high as target cell was chosen, and the effect of NECL1 on the proliferation of T98G cell line in vitro was detected by using cell growth curve, flow cytometry, and Hoechst staining.NECL1 was abrogated or markedly reduced in 6 glioma cell lines. When NECL1 was overexpressed in T98G cell line, the cell growth rate obviously decreased and the number of apoptotic cells remarkably increased when compared with the control group.NECL1 may inhibit the proliferation of T98G cells by inducing its apoptosis.

To study the role of cell adhesion molecules Necl1 in synaptogenesis in primary cultured neurons.Semi-quantitive reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression pattern of Necl1 in the neuronal differentiation cell model in vitro. Western blot was performed to detect the expression pattern of Necl1 in primary cultured rat neurons and in purified synaptosome. Immunofluoresence was used to detect the synapse formation in primary neurons and in 293 cells co-culture and to detect the density of synapses in primary neuron with ectopic expression of Necl1.Necl1 expression increased after retinoic acid (RA) induction in SH-SY5Y and P19 cells. The increase of Necl1 expression was consistent with the days of primary neurons culture in vitro, and Necl1 partly localized in synaptosome. The overexpression of Necl1 in 293 cells induced the synapse formation between cocultured 293 cells and neurons. Ectopic expression of Necl1 in primary neurons increased the density of synapses.Necl1 plays an important role in neuronal synapse formation.

To detect the expression of CBX7 in human glioma and investigate the potential regulatory effect of abnormally expressed microRNAs on CBX7 expression.Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot were applied to detect the expression pattern of CBX7 in 2 human normal brain tissues, 9 glioma tissues, and 3 glioma cell lines. Miranda algorithm and Ensemble Machine Learning algorithm were combined to predict miRNAs that target human CBX7. The expression of miR-9 in those tissues and cell lines were detected by real-time PCR. After miR-9 overexpression in 293ET and miR-9 knock-down in T98G, luciferase assay and Western blot were used to confirm the effect of miR-9 on CBX7 expression. MTT assay and flow cytometry were applied to detect the effect of miR-9 knock-down on T98G cells.No obvious difference in the CBX7 mRNA level between normal and tumor tissues was observed, while the protein level of CBX7 was abrogated or markedly reduced in glioma tissues and cell lines. Several miRNAs including miR-9 may target CBX7 by bioinformatics prediction. MiR-9 was up-regulated in glioma tissues and cell lines. In 293ET cell, luciferase activity of CBX7-3'UTR reporter was decreased to 24% after miR-9 overexpression. After miR-9 knock-down in T98G cell, the luciferase activity was increased by 1.8 fold and there was no change of CBX7 mRNA, while the protein level of endogenous CBX7 was significantly increased. The number of survival T98G cells increased and cells in G1 phase decreased after miR-9 knock-down.In human glioma, CBX7 is down-regulated by the inhibition of miR-9 at posttranscriptional level.

To generate a sensitive tool for noninvasive monitoring of a therapeutic gene vasostatin.We fused the bioluminescent reporter gene firefly luciferase to the therapeutic transgene vasostatin and ensured that these two proteins would not interrupt each other and kept their own natural character.We therefore examined clones of PC3 cells stably expressing fusion gene and positive controlfluc with bioluminescence. In vivo imaging of PC3-Fluc subcutaneous tumors showed that the mean tumor bioluminescence increased in animals over several weeks.Noninvasive monitoring facilitates the detection of gene expression in vivo and in vitro.

To express and purify the recombinant human pigment epithelium-derived factor (PEDF) which inhibits the proliferation of the endothelium cells from blood vessel in E.coli.PEDF gene was inserted into the prokaryotic expression vector pGEX-4T-2. The recombinant protein PEDF was expressed in E.coli BL-21, and purified by the GST Sepharose 4B affinity column. The recombinant human PEDF protein was identified by Western blot and mass spectrum. The biological activity of the recombinant human PEDF protein was measured by using MTT.The 46 kDa recombinant human PEDF protein was obtained. It significantly inhibited the proliferation of the human umbilical vein cell line HUVEC.The recombinant human PEDF with anti-angiogenesis activity protein may be successfully purify through prokaryotic expression.

To study the distribution of collagen IX gene in the disc and to determine its role in the pathogeny of idiopathic scoliosis (IS).The data included apical disc and intermediate disc from 14 cases of adolescent IS, 26 discs from 13 cases of scoliosis of confirmed pathogeny (CPS), which included 10 cases of congenital scoliosis and neurofibromatosis scoliosis. Six discs were obtained from 3 cases of normal young man served as controls. The distribution of collagen IX was studied in the apical disc of IS by immunohistochemistry and in situ hybridization (ISH) with RNA probe. The figure of collagen IX hybridization in the endplate cartilage was input to the figure analysis system. The mRNA content of collagen IX was compared between each group by SPSS software.Collagen IX was mainly distributed in the inner fibrous annulus, nucleus and endplate cartilage. Collagen IX was secreted by the little round chondrocyte-like cells, which was not expressed in the hypertrophic cells. There was significant difference of collagen IX mRNA content between the concave side of apical disc in the IS and the normal disc(P0.05), and also between intermediate vertebrae of CS group and normal.There is no obvious abnormal distribution of collagen IX in the disc of idiopathic scoliosis. Collagen IX may be related to the pathogensis of IS. More investigation such as quantity analysis and protein function determination is needed to confirm its role in the pathogenicity of IS.

Human DPY-30-like is a homolog of C. elegans DPY-30. DPY-30 is an essential component of dosage compensation machinery and loss of dpy-30 activity results in XX-specific lethality. In XO animals, DPY-30 is required for developmental processes other than dosage compensation. In yeast, the homolog of DPY-30, Saf19p, functions as a member of histone 3 lysine 4 methylation complex, which is the key part of epigenetic developmental control. In this report, human DPY-30-like protein was overexpressed and purified with the goal of structure determination. It was crystallized at 291 K in hanging drops by the vapour diffusion technique from a precipitant solution consisting of (NH4)2SO4 (1.5-2.0 M), Tris-HCl (0.1 M, pH 8.0). The crystal diffracted to 2.7 A resolution at 100 K in-house and belongs to the space group P4(1)2(1)2 or P4(3)2(1)2 with unit-cell parameters of a=b=74.5 A, c=87.0 A, alpha=beta=gamma=90.0 degrees. The asymmetric unit contains two molecules with 49% solvent content. We also analyzed its biochemical and biophysical characterizations. Efforts are now under way to determine the molecular structure of the DPY-30-like. These studies will open a new avenue towards the structure-based functional analysis of human DPY-30-like and dosage compensation machinery.

To study the distribution of collagen X in the intervertebral disc and determine its role in the pathogeny of idiopathic scoliosis (IS).The data included apical disc and intermediate disc from 14 cases of AIS, 26 discs from 13 cases of scoliosis of confirmed pathogeny which included 10 cases of congenital scoliosis and neurofibromatosis scoliosis (CS group). Six discs were obtained from 3 cases of sudden death of normal young man served as controls. The distribution of collagen X in the apical disc of IS was examined by immunohistochemistry and in situ hybridization (ISH) with RNA probe. The figure of collagen X hybridization in the endplate cartilage was input to the figure analysis system. The mRNA content of collagen X was compared between every 2 groups by SPSS software.Collagen X was mainly distributed around the hypertrophic chondrocyte in the endplate cartilage. Its mRNA was expressed in the hypertrophic chondrocyte. Positive signal was found in the nuclei of several scoliosis patients, which was not related to the pathogeny. The collagen X mRNA contents of the apical disc and intermediated disc of the IS group and of the CS group were significantly higher than those of the normal group (P0.05). There was no difference between the IS and CS groups.Collagen X is mainly distributed around the hypertrophic chondrocyte in the endplate cartilage. Its can also be secreted by the chondrocyte-like cells in the nucleus under special condition. Higher expression of collagn X gene in scoliosis patients may be the effects of long term abnormal stress which causes calcification of endplate cartilage. Collagen X expressed in the nucleus may be the result of secretion of chondrocyte-like cells in the disc under abnormal mechanical condition.

Responses of Sesbania rostrata and S. cannabina to Pb, Zn, Cu and Cd toxicities were assessed by a seed-suspending seedbed(SSS) approach. The results showed that the SSS approach was suitable for testing the tolerance of a plant to the stress of toxic metals. The endpoints include seed germination success, straightened radicle and hypocotyl of the seedlings from the seeds. The measurements could be done easily and accurately. It was found that the elongation of radicle was the most sensitive indicator to the stress of heavy metals among the endpoints. When exposure to lower or medium concentrations of Pb, Zn, and Cd, the development of the lateral roots were favorable. Species of S. rostrata was more tolerant than S. cannabina to the heavy metals, especially to Zn and Cd. The ED50 of Pb, Zn, Cu and Cd were 32.90, 5.32, 4.40 and 12.00 microg/ml for S. rostrata, respectively, and they were 30.11, 2.87, 4.05 and 4.94 microg/ml respectively for S. cannabina.

To clone, express and purify nucleocapsid protein from SARS coronavirus PUMC2 strain.According to the published SARS coronavirus genome sequences, the full length cDNA of N protein from SARS coronavirus PUMC2 strain was cloned by RT-PCR and the cDNA was cloned into the pET32a expression vector. The recombinant N protein was expressed in E. coli BL21 (DE3), and purified by Ni(2+)-NTA.Prokaryoticly expressed and purified N protein of SARS coronavirus PUMC2 strain was obtained.The SARS coronavirus recombinant N protein obtained by genetic engineering methods can be used for further functional study of SARS coronavirus N protein.

To screen out the specific antibody clones against amyloid beta peptide 40, and clone the antibody gene and express it in a bacterial system, so as to provide a solid basis for novel diagnostic and therapeutic methods for Alzheimer's Disease.beta amyloid peptide 40 was bound on the solid surface of Nunc plates as antigen to screen the binding clones from a phage-display human single-chain Fv antibody library. After five rounds of bio-panning, the host E. coli TG1 was infected with eluted filamentous phage from the last turn of selection. 55 well-separated colonies were picked randomly from the plates and the specific positive clones were identified by ELISA test. The single-chain Fv antibody gene was sequenced and their amino acids sequence was deduced. The scFv antibody gene was sub-cloned into a protokayotic expression vector pGEX-6P-1 and transformed into bacteria strain BL21 to express the glutathione-S-transferase (GST) fusion single-chain antibody.ELISA test showed that 33 of the 55 clones could bind amyloid beta peptide 40 and 10 of the 33 clones could be inhibited by amyloid beta peptide 40 itself to below 50% of its original binding activities. Five of the 10 clones could also be inhibited by amyloid beta peptide 1-16 to the same level, which meant that the binding epitope of the antibody from the 5 clones was between first to sixteenth amino acids at amino-end of amyloid beta peptide 40. DNA sequencing data demonstrated that the gene of the single-chain antibody specifically against amyloid beta peptide 40 was consisted of 768 bp and the deduced amino acids sequence confirmed its typical antibody structure. The complement determinant regions and framework regions were discriminated empirically. After cloning the antibody gene into a protokayotic system, the GST fusion antibody was expressed as the expected size.After five rounds of bio-panning and subsequently serial ELISA testing, the specific antibody clones against amyloid beta peptide 40 were screened out successfully. The antibody gene DNA sequence and amino acids sequence were analyzed and confirmed. The fusion antibody was expressed as expected in the bacterial system.

To get the cDNA clones which cover the whole genome of SARS-CoV PUMC2 strain.Using the SARS-CoV PUMC2 strain genomic RNA as the template, the cDNA fragments were amplified by RT-PCR, the PCR products were further purified and ligated into the pGEM-T vector, and all the clones obtained were sequenced.The cDNA clones which cover the whole genome of SARS-CoV PUMC2 strain were obtained.These cDNAs can be provided for the function study of SARS-CoV proteins and the construction of full-length infectious cDNA clone of SARS-CoV.

To identify and clone the gene encoding human TNFLP (tumor necrosis factor like protein) for some functional study on TNFLP.The full-length cDNA of TNFLP was isolated from fetal brain cDNA library. Several kinds of software were used to analyze nucleotide sequence and amino acid sequence of TNFLP. TNFLP mRNA distribution was identified by Northern blot. TNFLP-C and TNFLP-N were expressed in E. coli with GST expression system.The cDNA of human TNFLP was 2,112 bp, which encoded protein of 208 amino acid. Hydrophobility analysis found there were two hydrophobility regions of human TNFLP. TNFLP-C (112-207 amino acid) and mouse TNF-alpha were homologous. The identity of their amino acid sequence was 42%. Moreover, both of them had a motif-TYKRL. TNFLP was located in chromosome 16. Human TNFLP was widely expressed in various human tissues. Northern blot showed TNFLP was highly expressed in heart, brain and spleen, only one transcript can be seen. GST-TNFLP-C and GST-TNFLP-N fusion proteins were obtained.Tissue expression spectrum of TNFLP and prokaryotic expression of TNFLP have been done, which establish the base for the functional analysis of TNFLP.

To prokaryoticly express and purify HuD protein and its RNA recognition motifs.HuD protein was prokaryoticly expressed and purified by molecular cloning technology. Its biologic activity was testified by Western Blot.Purified HuD protein and its RNA recognized motifs were observed.The result might aid for basic research and clinical application.

To identify and clone the gene encoding human M96 gene and study its expression spectrum in several blood cell lines.According to the sequence of human EST which was highly homologous to the mouse M96 gene, primers used for library screening were synthesized, then the human adult testis and fetal brain cDNA library were screened. The gene was analyzed by making use of BLAST and CLUSTAL W, and its expression spectrum was studied by multiple-cell lines Northern blot analysis. The expression change of M961 in cell differentiation was observed by use of K562 cell line induced by hemin.Two cDNA clones encoding human M96 gene were isolated, identified and named as M961, and M962. They were found to be isoforms of each other. Northern, blot showed that M961 gene was expressed highly in CEM, Hel, Dami and K562 cell lines. However, during K562 cell line differentiation, process the expression of M961 elevated only slightly.M961 gene was expressed highly in pluripotent cell lines with erythrocytic and megakaryocytic potentials.

Human calcyclin binding protein (hCacyBP) gene was obtained by the screening of a human cDNA library. The full coding region of CacyBP was cloned into E.coli strain pET28, and then was expressed and purified through affinity chromatography. Rabbit anti-human CacyBP polyclonal antibody was obtained by immunizing rabbit with the purified human CacyBP. Western blots showed that it was expressed extensively in many tissues of mouse. The results of immunohistochemistrial staining showed that the location of CacyBP in BT325 cell line before and after differentiation changed from cytoplasm into nucleus and perinucleus cytoplasm.

Cellular RNAs in eukaryotes undergo extensive posttranscriptional modifications, but as yet only a few RNA-modifying enzymes have been identified and characterized. Here we report on the cloning of FTSJ2 , a novel human gene encoding a putative RNA methyltransferase. FTSJ2 shares significant sequence homology with FtsJ/RrmJ, a recently identified Escherichia coli 23S rRNA uridine-2′- O -methyltransferase. FTSJ2 belongs to a new family of evolutionarily conserved S -adenosylmethionine-binding proteins. The gene FTSJ2 is located on chromosome 7p22 between MAD1L1 and NUDT1 . It is 8 kb in length, spanning three exons. Northern blot analysis revealed that the FTSJ2 transcripts are abundant in skeletal muscle, placenta, and heart, as well as in cancer cells. Immunofluorescence staining demonstrated that FTSJ2 protein localizes to the nucleolus. Our results suggest that FTSJ2 is likely a nucleolar RNA methyltransferase involved in eukaryotic RNA processing and modification.

Host cell factor 1 (HCF-1) is a cellular transcriptional coactivator which coordinates the assembly of enhancer complex through direct interactions with viral and cellular trans-activators such as VP16, Oct-1, LZIP, and GA-binding protein. These interactions are mediated by the beta-propeller domain comprising the first 380 residues of HCF-1 with six kelch repeats. Here we describe the identification and characterization of a novel HCF-like kelch repeat protein, designated HCLP-1. HCLP-1 is a ubiquitously expressed nuclear protein which is composed almost entirely of a six-bladed beta-propeller. HCLP-1 selectively interacts with LZIP but not with VP16. The physical interaction between HCLP-1 and LZIP leads to the repression of the LZIP-dependent transcription. The HCLP-1-binding domain of LZIP maps to residues 109-315, which contain the bZIP DNA-binding motif. Electrophoretic mobility shift assay demonstrates that HCLP-1 indeed interferes with the binding of LZIP to its DNA target. Thus, HCLP-1 serves a transcriptional co-repressor function mediated through its inhibitory interaction with the LZIP transcription factor. Our findings suggest a new mechanism for transcriptional regulation by HCF-like proteins.

We have previously characterized transcription factor LZIP to be a growth suppressor targeted by hepatitis C virus oncoprotein. In search of proteins closely related to LZIP, we have identified a liver-enriched transcription factor CREB-H. LZIP and CREB-H represent a new subfamily of bZIP factors. CREB-H activates transcription by binding to cAMP responsive element, box B, and ATF6-binding element. Interestingly, CREB-H has a putative transmembrane (TM) domain and it localizes ambiently to the endoplasmic reticulum. Proteolytic cleavage that removes the TM domain leads to nuclear translocation and activation of CREB-H. CREB-H activates the promoter of hepatic gluconeogenic enzyme phosphoenolpyruvate carboxykinase. This activation can be further stimulated by cAMP and protein kinase A. CREB-H transcript is exclusively abundant in adult liver. In contrast, the expression of CREB-H mRNA is aberrantly reduced in hepatoma tissues and cells. The enforced expression of CREB-H suppresses the proliferation of cultured hepatoma cells. Taken together, our findings suggest that the liver-enriched bZIP transcription factor CREB-H is a growth suppressor that plays a role in hepatic physiology and pathology. © The Author 2005. Published by Oxford University Press. All rights reserved., published_or_final_version

Background The downstream of tyrosine kinase/docking protein (Dok) adaptor protein family has seven members, Dok1 to Dok7, that act as substrates of multiple receptor tyrosine kinase and non-receptor tyrosine kinase. The tropomyosin-related kinase (Trk) receptor family, which has three members (TrkA, TrkB and TrkC), are receptor tyrosine kinases that play pivotal roles in many stages of nervous system development, such as differentiation, migration, axon and dendrite projection and neuron patterning. Upon related neurotrophin growth factor stimulation, dimerisation and autophosphorylation of Trk receptors can occur, recruiting adaptor proteins to mediate signal transduction. Results In this report, by using yeast two-hybrid assays, glutathione S-transferase (GST) precipitation assays and coimmunoprecipitation (Co-IP) experiments, we demonstrate that Dok6 selectively binds to the NPQY motif of TrkC through its phosphotyrosine-binding (PTB) domain in a kinase activity-dependent manner. We further confirmed their interaction by coimmunoprecipitation and colocalisation in E18.5 mouse cortex neurons, which provided more in vivo evidence. Next, we demonstrated that Dok6 is involved in neurite outgrowth in mouse cortex neurons via the RNAi method. Knockdown of Dok6 decreased neurite outgrowth in cortical neurons upon neurotrophin 3 (NT-3) stimulation. Conclusions We conclude that Dok6 interacts with the NPQY motif of the TrkC receptor through its PTB domain in a kinase activity-dependent manner, and works as a novel substrate of the TrkC receptor involved in NT-3-mediated neurite outgrowth in mouse cortex neurons.