Figure 5.

The effects of overexpression, inhibitor and SiRNA of PDI and NADPH oxidases inhibitor on changed total Nox1 in VSMCs induced by SS and/or AGEs. (a, b) The quiescent cultured VSMCs with stable PDI overexpression were treated with SS, AGEs or both for 1 h and continually cultured for additional 24 h; (c, d) The quiescent cultured VSMCs pretreated with DMSO (NC) or PDI inhibitor bacitracin (PDI-I) for 1 h were treated by SS and/or AGEs for 1 h and continually cultured for additional 24 h; (e–g) the cultured VSMCs transfected with siRNA-PDI-A, and -B (SiR-PDI-A and -B), siRNA-control (SiR-C) and lipofectamine (Lip); (h, i) the quiescent cultured VSMCs pretreated with DMSO (NC) or NADPH oxidase inhibitor (apopcynin, 0.1 mM) for 1 h, and then treated by SS and/or AGEs for 10 min. All treated cells above were then harvested to detect total PDI and/or Nox1or ox-PDI by Western blot. Beta-actin was set as an internal control. (b, d, f, g and i) Statistical results of ratios of total NOX1, PDI or ox-PDI/beta-actin from (a, c, e and h) from three independent experiments, respectively. a, P<0.05 versus negative control (NC); b, P<0.05 versus AGEs or SS of the same group; c, P<0.05 versus cells without PDI overexpression or without inhibitors of each group by ANOVA as indicated by LSD test. Data are shown as means±SEM