TGF-β increased PIASy, E12, and α-SMA mRNA and proteins in MCs.

<p><b>(</b>A) Mouse MCs were cultured in the presence or absence of 1 ng/ml TGF-β for the indicated times. Gene expression of PIASy, E12 and α-SMA was examined by quantitative RT-PCR using mRNA of MCs treated or untreated with TGF-β. Means of four independent experiments are shown. The results are presented as the fold-increase or decrease compared with the values of 0 hours. <i>Open bars</i> denote no treatment, <i>closed bars</i> TGF-β treatment. (C) Mouse MCs were cultured in the presence or absence of 1 ng/ml TGF-β for the indicated times. Total cell lysates were examined by Western blot analysis using anti-PIASy, anti-E12, and anti-α-SMA antibodies. Representative data from four independent experiments is shown. (B<b>)</b> Optical densitometry of PIASy, E12 and α-SMA in Western blotting. The values of PIASy, E12 and α-SMA were normalized for that of GAPDH and compared with the values of 0 hours. Means of four independent experiments are shown. <i>Open bars</i> denote no treatment, <i>closed bars</i> TGF-β treatment. (D) TGF-β increased PIASy proteins in MCs in a dose-dependent manner. Mouse MCs were cultured with the indicated doses of TGF-β for 24 hours. (E) TGF-β increased E12 proteins in mesangial cells in a dose-dependent manner. Mouse MCs were cultured with the indicated doses of TGF-β for 24 hours. The values of PIASy and E12 were normalized for that of GAPDH and compared with the values of control vehicle. Means of four independent experiments are shown. *P<0.05, v.s. control.