Figure 2 Inhibition of Klotho in vitro promotes the progression of epithelial-to-mesenchymal transition (EMT) and renal fibrosis. We stimulated the HK-2 cells by TGF-β1 (5 ng/ml) at various time points (0–72 h). Western Blot assay and RT-PCR were performed to investigate the protein and mRNA expression levels of Klotho in HK-2 cells (A, C), as well as the expression of EMT-related biomarkers: Fibronectin (A, D), α-SMA (A, E), and E-Cadherin (A, F). The quantitative analysis of western blotting (B) and PCR (C-F) indicated the potential relationship between Klotho and the progression of EMT. *** P<0.001 compared with control group; **** P<0.0001 compared to control group. Each experiment was performed at least 3 times.