Fig. 7: EHMT2 regulates VSMC proliferation and migration via the negative regulation of GADD45G.

a Representative western blot analysis and quantification of PCNA and SM22α expression in VSMCs treated with si-Scr, si-EHMT2 and combined EHMT2/GADD45G knockdown (si-EHMT2 + si-GADD45G) (n = 5 in each group). b The proliferation of VSMCs (n = 4) was detected via immunostaining of EdU, and nuclei were stained with DAPI. Scale bar, 50 μm. c Representative images of wound-healing assays. Images were taken at 0 and 12 h after scratching monolayers in VSMCs treated with si-Scr, si-EHMT2 and combined EHMT2/GADD45G knockdown (si-EHMT2 + si-GADD45G) (n = 3 in each group). d Representative images and quantitative analysis of transwell migration assay in VSMCs treated with si-Scr, si-EHMT2 and combined EHMT2/GADD45G knockdown (si-EHMT2 + si-GADD45G) (n = 5 in each group). Scale bar, 400 µm. e Percentages of VSMCs in different phases of the cell cycle after treatment with si-Scr, si-EHMT2 and combined EHMT2/GADD45G knockdown (si-EHMT2 + si-GADD45G) (n = 5 in each group). f,g Representative western blot analysis (f) and quantification (g) of cyclin B1, cyclin D1, CDK2, CDK4 and p21 expression in VSMCs treated with si-Scr, si-EHMT2 and combined EHMT2/GADD45G knockdown (si-EHMT2 + si-GADD45G) (n = 4 in each group). h Immunofluorescence staining showed the expression of GADD45G and cyclin D1 in si-Scr, si-EHMT2 and combined EHMT2/GADD45G knockdown (si-EHMT2 + si-GADD45G) transfected VSMCs (n = 4 in each group). Nuclei were stained with DAPI. Scale bar, 100 µm. The intensity was normalized to that of DAPI. P values were calculated using a one-way ANOVA (a–h).