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. 1999 Feb 16;96(4):1415–1420. doi: 10.1073/pnas.96.4.1415

Figure 6.

Figure 6

Y845F mutant receptor retains its ability to phosphorylate SHC and activate MAPK. COS-7 cells were transfected with plasmid DNA encoding HA-SHC or Flag-ERK2 and either Y845F or wt EGFR, cultured for 2 days, serum starved overnight, and left untreated or treated with 100 ng/ml EGF for 10 min. Extracts were immunoprecipitated with 12CA5 anti-HA antibody or anti-Flag M2 affinity gel and resolved by SDS/PAGE. The amount of tyrosine phosphorylated HA-SHC (a) and Flag-ERK2 (b) was observed by Western immunoblot analysis. The amount of wt EGFR or Y845F EGFR expressed in the populations of cells was the same as that shown for Fig. 4.