Figure 9

Changes in protein expression during H₂O₂-induced conversion of ECs into myofibroblasts inhibited by transfection of an siRNA against TGF-β1 and TGF-β2. (a–l) ECs transfected with an siRNA against TGF-β1 (siTGFβ1) (a–f), an siRNA against TGF-β2 (siTGFβ2) (g–l) or a non-targeting siRNA (siCTRL) (a–l), and incubated in the absence (−) or presence (+) of 10 μM H₂O₂ for 72 h, and then protein expression was analyzed. (a, c, e, g, i, and k) Representative images from western blot experiments performed for detection of the endothelial marker VE-cadherin (VE-cad) (a, g), fibrotic marker α-SMA (c, i) and ECM proteins fibronectin (FN) (e, k). Panels (b, d, f, h, j, and f) show densitometric analyses from several experiments, as shown in (a, c, e, g, i and k), respectively. Protein levels were normalized against tubulin, and the data are expressed relative to the siCTRL-transfected cells condition in the absence of H₂O₂. Statistical differences were assessed by a one-way analysis of variance (ANOVA) (Kruskal–Wallis) followed by Dunn’s post hoc test. **: P<0.01 against the siCTRL-transfected cells condition in the absence of H₂O₂. Graph bars show the mean±s.d. (N=3–4).