Fig. 4: EGFR palmitoylation critically contributes to metastatic CRC cell stemness in NAFLD. | Cell Death Discovery

Fig. 4: EGFR palmitoylation critically contributes to metastatic CRC cell stemness in NAFLD.

From: Non-alcoholic fatty liver disease promotes liver metastasis of colorectal cancer via fatty acid synthase dependent EGFR palmitoylation

Fig. 4

A Representative images of IHC staining for EGFR of MC38 allografts in control and NAFLD mice. Scale bars, 50 μm. B The protein level of EGFR of HCT 116 xenografts in control mice and NAFLD mice by western blot assay (n = 5). C The protein level of EGFR in HCT 116 and SW 480 cells treated with or without BSA-PA/OA (PA 100 µM, OA 200 µM) at indicated time points by western blot assay. D The protein level of EGFR in HCT 116 and SW 480 cells treated with or without BSA-PA/OA at indicated doses for 36 h by western blot assay (-PA 0 µM, OA 0 µM; PA 50 µM, OA 100 µM; PA 100 µM, OA 200 µM; PA 200 µM, OA 400 µM, respectively). E The palmitoylation level of EGFR in HCT 116 and SW480 cells treated with or without BSA-PA/OA (PA 100 µM, OA 200 µM) for 24 h by acyl-biotin exchange assay. F The palmitoylation level of EGFR in HCT 116 xenografts in control mice and NAFLD mice by acyl-biotin exchange assay. G The palmitoylation level of EGFR in HCT 116 and SW480 cells treated with BSA-PA/OA (PA 100 µM, OA 200 µM) and/or 2-BP (50 µM) by acyl-biotin exchange assay. H The protein level of EGFR in HCT 116 and SW480 cells treated with BSA-PA/OA (PA 100 µM, OA 200 µM) and 2-BP at indicated concentrations for 24 h by western blot assay. I The palmitoylation level of EGFR in in control cells (EGFR-WT) and EGFR palmitoylation deficient mutant cells (EGFR-9CS) treated with BSA-PA/OA (PA 100 µM, OA 200 µM) by acyl-biotin exchange assay. J The protein level of CSC markers Oct4, Nanog and SOX2 in control cells (EGFR-WT) and EGFR palmitoylation deficient mutant cells (EGFR-9CS) treated with BSA-PA/OA (PA 100 µM, OA 200 µM) for 36 h by western blot assay. K Sphere formation assay on control cells (EGFR-WT) and EGFR palmitoylation deficient mutant cells (EGFR-9CS) treated with BSA-PA/OA (PA 100 µM, OA 200 µM). Representative photos of sphere in second passage were taken on day 10 after cells were seeded (scale bars, 100 μm), sphere numbers and diameters were determined and plotted (n = 3). Data are presented as mean ± SEM. Significance was determined by two-tailed unpaired Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. BSA bovine serum albumin, PA palmitic acid, OA oleic acid, 2-BP 2-bromopalmitate.

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