Fig. 6
From: Promotion of growth factor signaling as a critical function of β-catenin during HCC progression
Epidermal growth factor receptor (EGFR) signaling promotes hepatocellular carcinoma (HCC) cell survival. a Growth curve of triple knockout (TKO) HCC cells upon α-catenin KD with shαcat1–2 (n = 6). α-Catenin and β-catenin protein levels upon expression of shαcat1–2, as detected by immunoblot. b Immunoblot for E-cadherin and N-cadherin in TKO HCC cells infected with empty MigR1 (control (CTRL)) or MigR1 dominant-negative form of E-cadherin (DNE). White arrowhead indicates DN-Ecad. c Immunoprecipitation (IP) of immonoglobulin G (IgG) and β-catenin in TKO HCC cells expressing control (CTRL) or DNE. The expressions of β-catenin, E-cadherin, and DNE were detected by immunoblot. Double arrowheads indicate two migrating forms of E-cadherin. d IF for E-cadherin (green), β-catenin (red), and 4′,6-diamidino-2-phenylindole (DAPI (blue) in TKO HCC cells and HEPG2 cells expressing control (CTRL) or DNE. Scale bar = 25 μm. e Immunoblot for β-catenin in TKO HCC cells expressing control (CTRL) or DNE. Cells were treated with 25 μg/ml cycloheximide (CHX) for the time indicated (0–24 h). f Growth curve of TKO HCC cells upon EGFR KD with shEGFR(A–B) (n = 3). EGFR and EGFR2(HER2) protein levels upon expression of shEGFR(A–B), as detected by immunoblot. g Representative fluorescence-activated cell sorting (FACS) plot for AnnexinV-FITC expression on TKO HCC cells upon 0–50 μM Afatinib, Erlotinib, and Sorafenib treatment for 24 h. Dosage conditions that have significant effect on cells are highlighted in red. h Representative phase contrast images of TKO HCC organoids upon 0–50 μM Afatinib, Erlotinib, and Sorafenib treatment for 10 days. Dosage conditions that have significant effect on organoids are highlighted in red. Data are represented as mean ± SEM. ***p < 0.001. n.s. not significant. See also Supplementary Fig. 6
