Nephrin cell surface expression in podocytes. (A) Confocal microscopic analysis (×600) for the distribution of nephrin in HGEC. Cells were fixed and stained with anti-nephrin antibody and AlexaFluor 488-conjugated anti-goat IgG as secondary antibody. (B) FACS analysis of nephrin cell surface expression. Representative experiments from flow cytometric analysis of nephrin. Thick grey lines represent HGEC 5 mM, thin grey lines represent the corresponding isotype control antibody and black lines represent stimuli. (C) Percentage of positive cells was calculated for the histogram section indicated as M1 (**p<0.01 vs. HGEC:5 mM).