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Additional file 2: Table S1. Antibodies. Table S2. Sequences of mimic and inhibitor. Table S3. Primers for qRT-PCR.

Additional file 1: Figure S1. The efficiency of miR-325-3p inhibitor and mimic in chondrocytes and the effect of miR-325-3p inhibitor and mimic on Collagen II in vitro. (A) Quantitative PCR analysis of miR-325-3p in mouse chondrocytes treated with miR-325-3p inhibitor or NC-inhibitor. n = 3 per group. (B-C) Representative Collagen II (green) immunofluorescence staining and quantitative analysis in normal chondrocytes treated with miR-325-3p inhibitor or NC-inhibitor. n = 6 per group. Scale bar: 50 µm. (D) Quantitative PCR analysis of miR-325-3p in CTS-treated chondrocytes administrated with miR-325-3p mimic or NC-mimic. n = 3 per group. (E-F) Representative Collagen II (green) immunofluorescence staining and quantitative analysis in CTS-treated chondrocytes administrated with miR-325-3p mimic or NC-mimic. n = 6 per group. Scale bar: 50 µm. All data are shown as the mean ±standard deviation (SD). **P < 0.01. Figure S2. AAV-miR-325-3p OE alleviates catabolism and promotes anabolism of chondrocytes in vivo. (A) Representative immunohistochemistry images of Collagen II (top), Aggrecan (middle) and MMP13 (bottom) in LFJ cartilage in sham, AAV-NC-treated groups and AAV-miR-325-3p OE-treated groups after 10 weeks bipedal standing. (B-D) Quantitative analysis of Collagen II, Aggrecan and MMP13 in (A). Scale bar=50 μm. n=6 per group. All data are shown as the mean ±standard deviation (SD). **P < 0.01. Figure S3. The effect of NSC-207895 (p53 activator) on mechanical-stress chondrocytes after treated with miR-325-3p mimic. (A) Representative immunofluorescence images of Collagen II in mouse chondrocytes in different groups. (B) Quantitative analysis of Collagen II fluorescence intensity in (A). Scale bar=50 μm. n=6 per group. All data are shown as the mean ±standard deviation (SD). **P < 0.01. Figure S4. Symmetrical facet joint degeneration was observed in the bipedal standing mice model. (A-B) Representative histological images of left and right FJs (Facet Joints) with Hematoxylin-eosin (H&E) and Safranin O/Fast Green staining after 10 weeks standing in mice. (C) Quantitative analysis of Osteoarthritis Research Society International (OARSI) score in (B); Scale bar=50 μm. All data are shown as the mean ±standard deviation (SD). n=6 per group. All data are shown as the mean ±standard deviation (SD). ‘n.s.’ represents non-significant (two-tailed Student’s t-test).

Effective methods for visualizing neurovascular morphology are essential for understanding the normal spinal cord and the morphological alterations associated with diseases. However, ideal techniques for simultaneously imaging neurovascular structure in a broad region of a specimen are still lacking. In this study, we combined Golgi staining with angiography and synchrotron radiation micro-computed tomography (SRμCT) to visualize the 3D neurovascular network in the mouse spinal cord. Using our method, the 3D neurons, nerve fibers, and vasculature in a broad region could be visualized in the same image at cellular resolution without destructive sectioning. Besides, we found that the 3D morphology of neurons, nerve fiber tracts, and vasculature visualized by SRμCT were highly consistent with that visualized using the histological method. Moreover, the 3D neurovascular structure could be quantitatively evaluated by the combined methodology. The method shown here will be useful in fundamental neuroscience studies. Supplementary Information The online version contains supplementary material available at 10.1007/s12264-021-00715-7.