Figure 1: SKIP and TGF-β1-induced uPA, PAI-1, and MMP-9 expressions in PC-3 cells. The effect of SKIP knockdown on TGF-β1-induced Smad3 phosphorylation in PC-3 cells. (a) Expression of SKIP in PC-3 cells treated with TGF-β1 determined by Western blot and RT-PCR; α-tubulin and GADPH were used as a control of protein and cDNA loading, respectively. (b) Zymography analysis of uPA and MMP-9 in PC-3 cells treated with TGF-β1 for 24 h. (c) RT-PCR analysis for PAI-1 expression in PC-3 cells treated with TGF-β1 for 24 h. GADPH was used as housekeeper gene to verify the equal amount of cDNA in each sample. (d) SKIP siRNA knockdown in PC-3 cells determined by Western Blot. Cells were transfected with control siRNA (siR-C) or siRNA against SKIP (siR-SKIP). (e) Smad3 phosphorylation after 60 min of TGF-β1 treatment, determined by Western Blot in siR-C or siR-SKIP transfected cells.