Effect of siRNA specific for PIASy and E12 in α-SMA regulation in MCs. (A) PIASy mRNA and protein were reduced by transfection of siRNA against PIASy in mouse MCs. Gene expression of PIASy was examined by quantitative RT-PCR using mRNA of MCs transfected with siRNA against PIASy or control siRNA (20 nM). (B) α-SMA mRNA expression was increased by transfection of siRNA against PIASy. (C) Western blots of MCs transfected with siRNA against PIASy or control siRNA. (D and E) Optical densitometry of PIASy and α-SMA in immunoblotting. (F) E12 mRNA and protein were reduced by transfection of siRNA against E12 in mouse MCs. Gene expression of E12 was examined by quantitative RT-PCR using mRNA of MCs transfected with siRNA against E12 or control siRNA (10 nM). (G) α-SMA mRNA expression was suppressed by transfection of siRNA against E12. (H ) Western blot analysis of MCs transfected with siRNA against E12 or control siRNA. (I and J) Optical densitometry of E12 and α-SMA in immunoblotting. Means of four independent experiments are shown. The results were presented as the fold-increase or decrease compared with the values of cells transfected with control siRNA. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. *P