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. 2005 Feb 3;102(7):2328–2333. doi: 10.1073/pnas.0409675102

Fig. 3.

Fig. 3.

Association of SOCS-5 with the EGF-R complex. BaF/ERX cells stably expressing Flag-tagged SOCS-5 (S5), SOCS-5 lacking the N terminus (ΔNT), and SOCS-5 with a mutated SH2 domain (mSH2) or with a deleted SOCS box (ΔSB) were incubated with (+) or without (-) 50 ng/ml EGF for 15 min on ice and lysed. EGF-R proteins were immunoprecipitated by using monoclonal antibody 528, and associated Flag-tagged SOCS proteins were detected by Western analysis with rat anti-Flag antibody (A). Total cell lysates were analyzed by Western blot with anti-phosphotyrosine antibodies (B). Western blots were reprobed by using anti-EGF-R monoclonal antibody 806 (C). Levels of SOCS-5 protein were determined by immunoprecipitation with anti-Flag antibodies conjugated to agarose (M2-beads) and Western blot with rat anti-Flag antibodies (D).