For histological analysis, kidney tissues fixed with 4% (v/v) buffered paraformaldehyde were embedded in paraffin, and 4-μm-thick sections were prepared. The sections were then stained with HE (haematoxylin and eosin) and Masson's trichrome.
Immunofluorescence analysis was carried out using anti-ALR polyclonal antibody, anti-α-SMA monoclonal antibody (Santa Cruz Biotechnology) and anti-vimentin monoclonal antibody (AbCam). Cryostat sections (4-μm thick) were fixed in cold acetone, and then permeabilized with 0.4% (v/v) Triton X-100 for 14 min, blocked in 5% (v/v) goat serum for 60 min and then incubated with different primary antibody overnight at 4°C in refrigerator, followed by the incubation with FITC-labelled anti-rabbit IgG (immunoglobulin G) (Zhongshan Goldenbridge Biotechnology) for 90 min. The slides were visualized by confocal laser-scanning microscopy (TCS-SP2, Leica). In each experimental setting, immunofluorescent images were captured with identical light exposure time.
Immunofluorescence analysis was carried out using anti-ALR polyclonal antibody, anti-α-SMA monoclonal antibody (Santa Cruz Biotechnology) and anti-vimentin monoclonal antibody (AbCam). Cryostat sections (4-μm thick) were fixed in cold acetone, and then permeabilized with 0.4% (v/v) Triton X-100 for 14 min, blocked in 5% (v/v) goat serum for 60 min and then incubated with different primary antibody overnight at 4°C in refrigerator, followed by the incubation with FITC-labelled anti-rabbit IgG (immunoglobulin G) (Zhongshan Goldenbridge Biotechnology) for 90 min. The slides were visualized by confocal laser-scanning microscopy (TCS-SP2, Leica). In each experimental setting, immunofluorescent images were captured with identical light exposure time.