Signaling through Akt and MAPK but not p38 MAPK is maintained in the absence of EGFR tyrosine 845 phosphorylation. (A) MCF7 cells induced to express wt-EGFR or Y845F-EGFR or (B) SUM149 cells treated with dasatinib for 2 hours were treated with 10 ng/ml EGF for 15 minutes. Cell lysates were made, and subsequent separated protein was immunoblotted using pGab1-Tyr627, Gab1, pAkt-Ser473, Akt, pMAPK-Thr185/Tyr187, MAPK, pp38 MAPK-Thr180/Tyr182, and p38 MAPK antibodies. Each experiment was performed at least three independent times.