Fig. 7

p53 participated in ROCK1-mediated lens epithelial cells apoptosis in response to H₂O₂ treatment. a Western blotting confirmation of the siRNA-mediated p53 knockdown. Cells were lysed for western blotting after transfection with si-p53 and si-NC, respectively. β-actin was used as an internal reference. Data are based on three independent experiments. The 40–70 kDa blot was retained by horizontal shear and incubated with a p53 antibody. The 35–50 kDa blot was retained by horizontal shear and incubated with the β-actin antibody. b The knockdown of p53 resisted the decreased cell survival index after ROCK1 over-expression. Cells transfected with the pcDNA-ROCK1 and si-p53 and were tested by the MTT assay. **p < 0.01 versus the pcDNA-NC and si-NC group. ^##p < 0.01 versus pcDNA-ROCK1 and si-NC group, (n = 3). c The knockdown of p53 resisted the increase in apoptotic cells after ROCK1 over-expression. The apoptosis detection of LECs after transfection with pcDNA-ROCK1 and si-p53. Scale bar: 50 µm. d The apoptosis rates, which were calculated based on at least 100 cells from three experiments, **p < 0.01 versus the pcDNA-NC and si-NC group. ^##p < 0.01 versus pcDNA-ROCK1 and si-NC group. e Western blotting indicated that knockdown of p53 suppressed the increase in cleaved caspase3 levels by ROCK1 overexpression. The 10–30 kDa blot was retained by horizontal shear and incubated with a cleaved caspase3 antibody. The 35–50 kDa blot was retained by horizontal shear and incubated with the β-actin antibody. f Cells transfected with si-ROCK1 and p53-GFP were treated with H₂O₂ for 24 h and cell survival was tested by the MTT assay (n = 5). **p < 0.01 versus si-NC and GFP group. ^##p < 0.01 versus si-NC and GFP under H₂O₂-treated group. ^$$p < 0.01 versus si-ROCK1 and GFP under H₂O₂-treated group. Data are based on three independent experiments. g Cells transfected with si-ROCK1 and p53-GFP were treated with H₂O₂ for 24 h and cell apoptosis was visualized by the Hoechst 33342 staining assay. h Cell apoptosis was analyzed after transfection with si-ROCK1 and p53-GFP plasmids. **p < 0.01 versus si-NC and GFP group. ^##p < 0.01 versus si-NC and GFP under H₂O₂-treated group. ^$$p < 0.01 versus si-ROCK1 and GFP under H₂O₂-treated group. Scale bar: 50 µm