Fig. 3.

Myo1c is a podocyte protein and colocalizes with Neph1. (A) Kidney sections from PFA-perfused rats were immunostained with Myo1c and Neph1 antibodies to determine the colocalization of Myo1c (red) with Neph1 (green) in glomeruli. (B) To determine the specificity of the Myo1c antibody, it was preincubated with the Myo1c protein prior to staining. Nuclear staining was performed with DAPI (blue). (C and D) Immunogold electron micrographs of kidney sections stained with Myo1c antibody show Myo1c localization adjacent to the slit diaphragm (arrows). P, podocytes; GBM, glomerular basement membrane; SD, slit diaphragm. (E and F) Lysate obtained from mouse glomeruli (E) or cultured human podocytes (F) was subjected to flotation gradient centrifugation using Optiprep. Fractions were analyzed by immunoblotting with the indicated antibodies. Western blotting with caveolin antibody identified the lipid raft fraction (fraction 3 at the interface between the 5 and 30% Optiprep densities).