Figure 6

Silencing of InsR disrupted InsR/IGF-1R hybrid receptor formation and increased IGF-1R homodimer formation. (A) Lysates from non-transfected MES-13 mesangial cells were immunoprecipitaed (IP) using antibodies against IGF-1Rα, InsRα or control (CTR) IgG, an irrelevant control antibody. Immunoprecipitates were immuneblotted (IB) for IGF-1Rβ or InsRβ. WCL, whole cell lysate. An image of gel stained after transfer was shown as a loading monitor (lower panel). (B) Hybrid receptors in SC or InsR shRNA (sh-InsR) transfected cells were detected by immunoprecipitation with antibodies directed against either the InsRα or IGF-1Rα. Immunoprecipitated receptors were detected by immunoblotting with an anti-IGF-1Rβ antibody (upper panel). An image of gel stained after transfer was shown as a loading monitor. Immunodepletion was assessed by immunoblotting the supernatants from immunoprecipitate with an anti-IGF-1Rβ antibody (middle panel). The bar graph shows the densitometric scanning results (upper panel, means ± SEM) of four independents experiments *, P<0.05.