Quantification of Glomerular Proteins in ADR and MCD

The isolation and homogenates of the rat glomeruli and western blot analysis were performed as previously described[11 (link)]. The volume of centrifuged urine from the ADR rats and MCD patients was 20 μl, and the quantity of lysates from the ADR rats was 100 μg. For the detection of Angptl4, synaptopodin and desmin, the blots were incubated with goat anti-rat Angptl4 (Santa Cruz Biotech, Santa Cruz, CA, USA, 1:200), mouse anti-rat synaptopodin (1:50, Progen, Heidelberg, Germany) or mouse anti-rat desmin (1:100, Abcam, New Territories, Hong Kong). The following secondary antibodies were used: horseradish peroxidase-conjugated rabbit anti-goat IgG (Jackson ImmunoResearch, West Grove, PA, USA, 1:5000) or horseradish peroxidase-conjugated goat anti-mouse IgG (Jackson ImmunoResearch, West Grove, PA, USA, 1:5000). All western blot results were normalized to β-actin.