Figure 6
Expression of Smurf2 and Smad7 in HPMCs following exposure to TGF-β with/without treatment of LY294002 and Akt-DN. Panel A: Bar graph represents the summary of Smurf2 mRNA expression, as detected by Real-time PCR analyses. Panel B: Represents of Smurf2 and Smad7 expression profiles following treatment of LY294002 or transfected with Akt-DN, as assessed by Western blot analyses. The β-actin served as a loading control. Panel C: The bar graphs represent the relative bands density of Smad7 (Panel C1) and Smurf2 (Panel C2) of Western blots. A decreased expression of Smad7 was observed in HPMCs incubated with TGF-β1, while the effect was abolished by the pre-treatment with LY294002 and transfection with Akt-DN, in addition, TGF-β1 significantly up-regulated the expression of Smurf2, which was inhibited with Akt-DN or LY294002. Panel D: Immunofluorescence microscopy revealed that TGF-β1 significantly increased Smurf2 expression in HPMCs, which was dramatically decreased by LY294002 (next to the extreme right panel), but only partially attenuated by transfection with Akt-DN (extreme right panel). Values are the mean ± SEM, n = 5, * p <0.01 vs control, # p <0.01 vs TGF-β1.