Hirudin treatment abolishes podocyte protection in diabetic FVL mice. (A) Bar graph summarizing frequency of apoptotic glomerular cells. (B) Expression of apoptosis regulators in renal cortex tissue samples of diabetic mice without or with hirudin treatment. Bar graph (top) and representative immunoblot (bottom) showing p53 and Bax expression (n ≥ 7 each group). (C) Podocyte number in glomeruli of diabetic mice without or with hirudin treatment as determined by WT-1 immunohistochemical staining. (D) Bar graph (top, data normalized for podocyte numbers) and representative immunoblot (bottom) showing GLEPP1 expression in renal cortex tissue samples of diabetic mice without or with hirudin treatment. Actin immunoblot is shown as loading control (n ≥ 7 each group). (E) Bar graph (top) and representative immunoblots of immunoprecipates (bottom) showing nephrin tyrosine phosphorylation in renal cortex tissue extracts of diabetic mice without or with hirudin treatment (n ≥ 7 each group). DM indicates diabetic mice (black bars); DM + Hir, diabetic mice with hirudin treatment (gray bars); *P < .05; **P < .01 (t test).