Nephrin expression in podocytes. (A) Western blot analysis for nephrin expression in HGEC. Total protein was extracted from cells cultured either in the presence of 5 mM (control) (lane 1) or 25 mM (lane 5) glucose. Lanes 2, 3 and 4 represent HGEC:5 mM-to-25 mM/1w, HGEC:5 mM-to-25 mM/2w and HGEC:5 mM-to-25 mM/4w respectively. Lane 6 represents HGEC:25 mM-to-5 mM/18w. Lanes 7 and 8 represent HGEC:25 mM/6w-to-5 mM/4w and HGEC:25 mM/18w-to-5 mM/4w respectively. Blots were reprobed with anti-tubulin antibody, to verify protein loads, against which the data were normalized. (B) Results were expressed as the mean ± SD of four independent experiments (*p<0.05 vs. HGEC:5 mM).