Fig. 3. Nox-dependent signalling regulates TGFBR1/Alk-5 mediated Smad2 linker region phosphorylation in human VSMCs.

a VSMCs were treated with TGF-β1 (2 ng/ml) for 30 min in the presence and absence of the TGFBR1 antagonist, SB431542 (SB) (10 μM) and the Nox inhibitor, DPI (1–20 μM) b VSMCs were treated with TGF-β1 (2 ng/ml) for 30 min in the presence and absence of the Nox inhibitor, apocynin (1 and 10 μM). Membranes were incubated with anti-phospho-Smad2 (Ser245/250/255) (1:1000) followed with peroxidase labeled anti-rabbit IgG (1:10000) and ECL detection. Anti-GAPDH was as loading control. Normalised data in each case are shown as mean ± SEM from three independent experiments and statistical significance was determined by One-way ANOVA followed by least significant difference post-hoc analysis. ^*p < 0.05 and ^**p < 0.01 compared with untreated control, ^#p < 0.05 and ^##p < 0.01 compared with TGF-β1