Immunofluorescence Staining of Kidney Markers

Immunofluorescence staining was performed on paraffin sections. The tissue sections were processed for the primary antibody incubation [31 (link)]. The primary antibodies against nephrin (1 : 500, Abcam, Cambridge, UK), WT1 (1 : 50, Abcam, Cambridge, UK), and AQP1 (1 : 200, Abcam, Cambridge, UK) were incubated at 37°C for 1 h, followed by the incubation with secondary antibodies (goat anti-rabbit lgG, RS23220, ImmunoWay, USA) for 1 h at 37°C. The sections were counterstained with DAPI (D9542, Sigma, USA) to visualize the nuclei and observed using a digital pathology scanner (VS120–S6–W, OLYMPUS).