Fig. 5: Palmitoylation promotes EGFR stabilization and plasma membrane localization in NAFLD liver metastatic CRC cells.

A HCT 116 and SW 480 cells treated with or without BSA-PA/OA (PA 100 µM, OA 200 µM) were incubated with cycloheximide (CHX) (25 μg/ml) and analyzed by western blot at the indicated time points. Right, quantification of the results (n = 3). B HCT 116 and SW480 cells treated with or without BSA-PA/OA (PA 100 µM, OA 200 µM) were incubated with EGF (25 µg/ml) and analyzed by western blot at the indicated time points. Right, quantification of the results (n = 3). C HCT 116 and SW 480 cells treated with or without BSA-PA/OA (PA 100 µM, OA 200 µM) were incubated with 25 μg/ml CHX and 5 µM MG132 and analyzed by western blot at the indicated time points. Right, quantification of the results (n = 3). D HCT 116 and SW 480 cells treated with or without BSA-PA/OA (PA 100 µM, OA 200 µM) were incubated with 25 μg/ml CHX and 250 µM NH₄Cl and analyzed by western blot at the indicated time points. Right, quantification of the results (n = 3). E Representative immunofluorescence images for EGFR (red), LAMP1 (green), and DAPI (blue) of HCT 116 cells treated with or without BSA-PA/OA (PA 100 µM, OA 200 µM). Scale bars, 5 μm. Intensity profiles of LAMP1 (green lines) and EGFR (red lines) co-localization signal were shown in plotted lines at three random sites. F Representative immunofluorescence images for EGFR (red), F-actin (green), and DAPI (blue) of HCT 116 cells treated with or without BSA-PA/OA (PA 100 µM, OA 200 µM). Scale bars, 10 μm. Data are presented as mean ± SEM. NAFLD non-alcoholic fatty liver disease, BSA bovine serum albumin, PA palmitic acid, OA oleic acid, MFI mean fluorescence intensity, 2-BP 2-bromopalmitate, FASN fatty acid synthases, CHX cycloheximide.