Figure 7
Role of Smurf2 and USP4 in TGF-β 1 or Akt-mediated ZO-1, Vimentin and Fibronectin expression in HPMCs. Panels A & B: The bar graphs represent the summary of the mRNA expression of ZO-1 and Vimentin, as detected by Real-Time PCR. Panel C: Western blot analyses indicated a decreased protein expression of ZO-1 in HPMCs treated with TGF-β1 or co-transfected with Flag-Akt. There was no obvious change in cells treated with Smurf2-siRNA and USP4 siRNA compared to basal levels. The TGF-β1-mediated reduced expression of ZO-1 was partially reversed with the transfection of Smurf2-siRNA and USP4 siRNA. ZO-1 expression was markedly decreased by the treatment with TGF-β1 and transfection with Flag-Akt. The Smurf2-siRNA and USP4 siRNA also partially restored the TGF-β1 + Flag-AKT reduced ZO-1 mRNA expression. Panel D, D1-D2: The bar graphs represent the Western blot relative band density of ZO-1 and Vimentin compared to β-actin. Panel E: Bar graphs represent the fibronectin concentration in the supernatant of cultured HPMCs, as detected by ELISA. Values are the mean ± SEM, n = 4, *p < 0.01 vs control. # p <0.01 vs TGF-β1, $ p <0.01 vs TGF-β1+ Flag-Akt.