Figure 2.

Neither TAK1 nor TRAF6 mediates TGFβ-induced ATF2 activation. VSMCs were transfected with 20 nM TAK1 siRNA (A) or TRAF6 siRNA (B) using Lipofectamine RNAiMAX transfection reagent and then stimulated with or without TGFβ for 10 min. Control siRNA was also transfected as a control. Western blot analysis was performed to detect TAK1, TRAF6, and phosphorylation of ATF2 and Smad2. The membranes were subsequently probed with actin for loading control. (C) VSMCs were transfected with control or TRAF6 siRNA and then treated with or without TGFβ for 24 h. Total proteins were then isolated for Western blot analysis to detect TRAF6 and CRP2 expression. The membranes were subsequently probed with actin for loading control. Representative blots of at least three independent experiments are shown.