Figure 4

ICR downregulation attenuates fibrotic changes and inhibits phosphorylation of Akt and mTOR in TGF-β1-treated human proximal tubular HK-2 cells. (a) QPCR analysis normalized to β-actin showed a significant decrease in the level of ICR in HK-2 cells transfected with ICR shRNA plasmids (sh-ICR) compared with its negative control (sh-ICR NC). Knockdown of ICR upon transfection with sh-ICR plasmid significantly reduced both profibrotic protein levels of collagen I and α-SMA (b), and phosphorylation of Akt and mTOR (c) in HK-2 cells stimulated with TGF-β1 (10 ng/ml, 48 h). The protein levels of collagen I and α-SMA were normalized to β-actin; the phosphorylated Akt and mTOR were normalized to Akt and mTOR, respectively. The original blots are presented in Supplementary Figs. S2 and Fig. S3. Data are presented as means ± SD of three independent experiments. *P < 0.05, significantly different from negative control group (sh-ICR NC or TGF-β1 + sh-ICR NC).