Fig. 4

EE02 blocks the EGFR/EPS8 interaction. a The schematics of the plasmids used in the study encoding wild-type EGFR and mutant-type EGFR are shown. To create the mut-EGFR plasmid, the oligonucleotides encoding the EGFR JXM domain (amino acids 650–679) were replaced by an unstructured sequence encoding ten GGS repeats as described in the Methods section. b Cancer cell lines transfected with wt-WGFR, mut-EGFR, EGFR/shRNA1, EGFR/shRNA2 and NC shRNA were subjected to western bolt analysis for EGFR and GAPDH expression. c Cells were untreated or treated with EE02 or DMSO for 24 h, then the cell lysates were immunoprecipitated with an anti-EGFR antibody or an anti-IgG antibody and were immunoblotted with the indicated antibody. d Cancer cell lines transfected with wt-WGFR, mut-EGFR, EGFR/shRNA1, EGFR/shRNA2 and NC shRNA were treated with EE02 for 24 h, 48 h or 72 h at increasing concentrations, and then cell viability was assessed using CCK-8 assays (n = 3)