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. 2015 Sep 12;6(33):34992–35003. doi: 10.18632/oncotarget.5252

Figure 4. C797 site is conserved in kinase active ErbB members and is important for constitutive activation and palmitoylation of EGFR.

Figure 4

a. Comparison of palmitoylation site (C797) of EGFR with HER2, HER3 and HER4. b. Western blotting analysis for palmitoylated EGFR prepared from HEK293 cells transfected with EGFR WT or C797G mutant (Flag-tagged). c. C797G mutation blocks EGFR constitutive phosphorylation when expressed in HEK 293 cells. d. C797G mutation blocks palmitoylation of ΔECD EGFR in HEK 293 cells. e. C797G mutation blocks ΔECD EGFR constitutive phosphorylation in HEK 293 cells. f. HEK 293 cells were transfected with WT or C797G EGFR for 24 hours and treated with DMSO or EGF at 10 ng/ml for 20 minutes and protein samples were analyzed for EGFR dimers using Western blotting under reducing or non-reducing conditions. g. The same samples from panel f were subjected to Western blot analysis. h. Western blot analysis for Flag, pErk 1/2, Erk 1/2 and GAPDH. Protein samples were prepared from HEK 293 cells transfected with WT or cysteine mutants of ErbB members.