Fig. 6.

miR-325-3p targeted to P53 and NSC-207895 (P53 activator) reversed miR-325-3p effect on suppressing mechanical stress-induced cellular senescence in vitro. A miR-325-3p targeted to multiple genes by TargetScan, miRDB databases, and cellular senescence GO databases (GO:0,090,398). B Quantitative PCR analysis of miR-325-3p targeted five genes in mouse chondrocytes treated with miR-325-3p mimic or NC-mimic. n = 3 per group. C Complementary sequences between miR-325-3p and the 3′ UTR of Trp53. D Relative luciferase activities of the p53-wt + negative control group, p53-wt + miR-325-3p group, p53-mut + negative control group, and p53-mut + miR-325-3p group. n = 3 per group. E Representative p21 immunofluorescence staining and SA-β-Gal images in normal chondrocytes and CTS (20% elongation, 24 h)-treated chondrocytes treated with miR-325-3p control or mimic or mimic plus NSC-207895. Red arrows represent SA-β-Gal.⁺ cells. Scale bar: 20 μm in the left SA β-Gal staining panel and 50 μm in the right IF panel. F, G Quantification of SA-β-Gal staining and P21 immunofluorescence staining in E. n = 6 per time point. H Quantitative PCR analysis of cellular senescence key genes (P16, P21, P53) and SASP-related genes (PAI-1, IL-6, TGF-β) in chondrocytes described in E. n = 3 per group. I, J Representative western blot images and quantification in chondrocytes described in E. n = 3 per group. All data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01