Protein Isolation and Western Blot Analysis

Total proteins were isolated from kidney and HK-2 cells and quantified as described previously [53 (link)]. Protein samples (30 μg/lane) were resolved using sodium dodecyl sulfate–polyacrylamide electrophoresis and then transferred onto polyvinylidene difluoride membranes (PVDF, Millipore, USA). The membranes were incubated at 4 °C overnight with α-SMA (1:300, Abcam, Cambridge, MA, USA), fibronectin (1:300, Santa Cruz, CA, USA), vimentin (1:500, Santa Cruz, CA, USA), E-cadherin (1:1000, Proteintech, Wuhan, China), Megalin (1:100, Santa Cruz, CA, USA) and HBc (1:1000, Abcam, Cambridge, MA, USA), followed by incubation with horseradish peroxidase-conjugated goat anti-mouse (1:10,000, Santa Cruz, CA, USA) or goat anti-rabbit IgG secondary antibodies (1:10,000, Santa Cruz, CA, USA) at room temperature for 1 h. The immunoblots were imaged by the chemiluminescence western blot detection system (Bio-Rad ChemiDoc MP, California, USA) with GAPDH (1:5000, ZENBIO, Chengdu, China) as the loading control.