Fig. 2: EGF promotes SHCBP1 nuclear translocation. | Cell Death & Disease

Fig. 2: EGF promotes SHCBP1 nuclear translocation.

From: EGF-induced nuclear translocation of SHCBP1 promotes bladder cancer progression through inhibiting RACGAP1-mediated RAC1 inactivation

Fig. 2

A Immunoblotting analysis showing total expression of SHCBP1 protein after EGF treatment (100 ng/ml) at the indicated time points in 5637 and T24 cells. B Immunoblotting analysis showing SHCBP1 redistribution in the cytoplasm and nucleus following EGF stimulation for 30 min in 5637, T24, and UMUC-3 cells. Quantified value of SHCBP1 compared to internal control was shown. C IF images indicating SHCBP1 localization from the cytoplasm to the nucleus after EGF treatment in 5637, T24, and UMUC-3 cells. D Immunoblotting analysis showing the expression of p-EGFR, p-AKT, and p-ERK1/2 in response to EGF at the indicated time points in 5637 and T24 cells. E Immunoblotting analysis determining the impact of a PI3K/Akt inhibitor (LY294002, 20 µM) or an ERK1/2 inhibitor (U0126, 10 µM) on the nuclear translocation of SHCBP1 in 5637 and T24 cells. F EGF-induced SHCBP1 nuclear translocation was examined after erlotinib (EGFR tyrosine kinase inhibitor) treatment in 5637 and T24 cells. G SHCBP1 nuclear translocation was examined by immunoblotting in the absence or presence of erlotinib in UMUC-3 cells. H SHCBP1 expression in bladder cancer patients with or without EGFR mutation was compared (data from TCGA). I Following immunoprecipitation against HA, SHCBP1 phosphorylation was detected using immunoblotting with an anti-phosphoserine (pSer) antibody in lysates from T24 cells transfected with HA-SHCBP1 (WT) or mutant HA-SHCBP1 (S273A) and treated with EGF. J, K Immunoblotting and IF showing the subcellular localization of HA-SHCBP1 or mutant HA-SHCBP1 (S273A) in T24 cells in response to EGF.

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