Anti collagen 4

For each mouse, quantitative analysis of glomerular volume, fractional mesangial area, and tuft area in paraffin-embedded kidney sections stained with PAS reagent was performed as described previously [33 (link)]. To examine collagen matrix, paraffin-embedded sections were stained with a picrosirius red stain [33 (link)]. Oil Red O staining was performed to evaluate lipid accumulation in frozen kidney tissues as described previously [33 (link)]. For immunohistochemistry, we used anti-nephrin (1:100; Progen biotechnik GmbH, Heidelberg, Germany), anti-F4/80 (1:200; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), anti-αSMA (1:200), and anti-collagen IV (1:200; Southern Biotechnology Associates, Birmingham, AL, USA) antibodies. Images were captured using a Zeiss microscope equipped with an Axio Cam HRC digital camera and Axio Cam software (Carl Zeiss, Thornwood, NY, USA). Staining intensities were then quantified using Image-Pro Plus 4.5 software (Media 149 Cybernetics, Silver Springs, MD, USA) as described previously [33 (link)]. Imaging for DAPI (1:1000), anti-PMP70 (1:200, Abcam, Cambridge, MA), and anti-catalase (1:200, Santa Cruz Biotechnology) antibodies was conducted using a confocal microscope (Carl Zeiss, Gottingen, Germany).