Fig. 2

SMYD5 modulates FLS proliferation response to IL-1β. A-C SMYD5 was knocked down using SMYD5 siRNA in IL-1β (10 ng/ml, 24 h) induced FLS, with control siRNA (siNC) as a negative control. A Immunoblot analysis and quantification of SMYD5, Cyclin D1, and MMP9, with β-tubulin used as a loading control, n = 6. B Smyd5 and Ccnd1 mRNA levels were detected by RT-qPCR analysis, n = 3. C Representative images of EdU staining in IL-1β-induced FLS pre-transfected with control or SMYD5 siRNA. Scale bars, 50 μm. Right panel shows statistical analysis of the proportion of EdU-positive cells, n = 6. D–F FLS were infected with lentivirus vectors encoding SMYD5 (LV-SMYD5) to promote SMYD5 overexpression. D Immunoblot analysis and quantification of SMYD5, n = 4. E Immunoblot analysis and quantification of Cyclin D1 and MMP9, with β-tubulin used as a loading control, n = 6. F RT-qPCR analysis of Smyd5 and Ccnd1 mRNA levels, n = 3. G Representative images of EdU staining in LV-SMYD5-treated or untreated FLS. Scale bars, 100 μm. Right panel shows the quantification of EdU-positive cells, n = 6. Data presented as mean ± SEM, p values calculated by two-tailed Student’s t-test (D–G) or one-way ANOVA test (A–C), ^***p < 0.001