Figure 5. Quantitative real-time PCR and immunoblotting showing the effect of vorinostat (2.5 µM), TGIF1, and TGIF2 siRNA knockdown on αSMA protein levels. Control human corneal fibroblast cultures (–TGFβ) showed no αSMA expression. Cultures treated with TGFβ1 (5 ng/ml) showed robust αSMA increase. Vorinostat (+TGFβ+vorinostat) treatment markedly attenuated TGFβ-evoked αSMA mRNA (A) and protein (B) expression. The knockdown of TGIF1 (+TGFβ+vorinostat+TGIF1siRNA) and TGIF2 alone (+TGFβ+vorinostat+TGIF2siRNA) or in combination (+TGFβ+vorinostat+TGIF1+2siRNA) significantly reversed the effect of vorinostat on αSMA. * p<0.001 compared to –TGFβ, Ψ p<0.001 compared to +TGFβ, # p<0.05 compared to +TGFβ+vorinostat.