Figure 2From: A PKC-SHP1 signaling axis desensitizes Fcγ receptor signaling by reducing the tyrosine phosphorylation of CBL and regulates FcγR mediated phagocytosisPMA treatment reduced the tyrosine phosphorylation of CBL and dissociation of binding of CRKL to CBL. Immunoprecipitations of U937 cell lysates were performed with anti-CBL antibody after PMA or FcγRI stimulation. Lane 1 & 2 indicates preimmune and resting state (NS) respectively. Lane 3, 4 & 5, treated with PMA (200 ng/ml) for 1, 5 and 10 min. respectively. Lane 6, 7 & 8, stimulated for FcγR1 with 32.2 F(ab)2 and cross linked with secondary antibody (rabbit anti mouse F(ab’)2) for 1 , 5 and 10 min. respectively. Whole cell lysate was added in lane 9 as positive control (WCL). Immunoprecipitated proteins were resolved in 10% SDS-PAGE, blot on nitrocellulose membrane and probed with anti-phosphotyrosine antibody (4G10). This blot was again reprobed with anti-CBL and anti-CRKL antibody. This experiment was repeated three times.Back to article page