Disruption of PC/NHERF2/ezrin complexes in glomeruli from PAN-, SA-, and PS-treated rats. (a) Immunoprecipitation with anti-PC followed by immunoblotting for NHERF2 and ezrin as for Figure 6. The amount of ezrin and NHERF2 that coprecipitates with PC is reduced significantly in SA-treated (lane 5) and PS-treated (lane 6), but not in PAN-treated, glomeruli (lanes 3 and 4) compared with normal controls (lane 2). The amount of ezrin and NHERF2 that coprecipitates is restored to normal after heparin (Hep) perfusion of PS-treated rats (lane 7). Lane 1: 10 μg glomerular lysate. (b) When immunoprecipitation is carried out with anti-ezrin followed by immunoblotting for NHERF2 and PC, a similar reduction is seen in the amount of PC that coprecipitates with ezrin in SA (lane 5) and PS (lane 6) treated rats but not in PAN nephrotic rats (lanes 3 and 4). No change is seen in the amount of NHERF2 coprecipitated (bottom panel). Glomerular lysates from normal, PAN 3-d, and PAN10d, SA- and PS-treated rats were immunoprecipitated with anti-PC (1A + 5A) mAb (a) or polyclonal anti-ezrin (b) followed by immunoblotting as in Figure 6. Lane 1: 10 μg control lysate. Arrowheads indicate desialylated PC, which has a slower mobility than intact PC (lane 5). The arrow indicates IgG heavy chain. The experiment was performed twice with comparable results.