Fig. 2
SHCBP1 translocates to the nucleus in response to EGF stimulation. a Following immunoprecipitation against SHC1, western blotting analysis reveals interaction between SHCBP1 and SHC1 in response to EGF treatment in the absence or presence of EGFR depletion. 293T and A549 cells were incubated with EGF (100 ng/ml) for 30 min. b WB analysis showing SHCBP1 redistribution in the cytoplasm and nucleus in response to EGF treatment. c, d WB examining nuclear SHCBP1 after EGF treatment at indicated concentrations (c) and time points (d) in 293T and A549 nuclear extracts. e Total SHCBP1 in whole-cell lysates were detected at indicated time points upon EGF treatment. f SHCBP1 expression in NSCLC patients with or without EGFR mutation was analyzed (data from TCGA). g WB results showing nuclear translocation of SHCBP1 stimulated by EGF in HepG2 liver cancer, MCF-7 breast cancer, and KYSE410 esophageal carcinoma cell lines. h Effects of SHC1 depletion on relocation of SHCBP1 to the nucleus. i WB analysis determining the impact of Ras/Erk inhibitor (U0126, 10 µM) or PI3K/Akt inhibitor (LY294002, 20 µM) on nuclear translocation of SHCBP1 in 293T cells. j SHCBP1 nuclear translocation was analyzed by WB assay in the absence or presence of EGFR tyrosine kinase inhibitor gefitinib in two NSCLC cell lines with EGFR activating mutations, HCC827 and HCC4006
