Fig. 2.

Western blot analysis of podocyte- and endothelial glomerular cell-specific proteins in cultured mesangial cells. A: expression of podocyte-specific proteins including podocin, CD2AP, and nephrin in podocytes and mesangial cells. Actin loading control is shown for podocin and CD2AP. Nephrin blotting was performed using nephrin immunopreciptation (black line indicates where irrelevant lanes have been removed). B: expression of VE-cadherin, an endothelial marker, in mesangial and endothelial cells. Actin was used as a loading control in A and B, and whole glomeruli as a positive control for all proteins except for CD2AP, where we used HEK cells as a positive control. Podocytes and not mesangial cells expressed CD2AP, podocin, and nephrin. VE-cadherin is expressed only in endothelial cells and not in mesangial cells. These results validate the staining results and provide further evidence of the establishment of a mesangial-specific cell line in culture.