Fig. 5

24(R/S),25-EC and 27HC regulate gastric epithelial function. a The levels of 8 oxysterols (25HC, 27HC, 24(S) HC, 24(R/S),25-EC, 7αHC, 7βHC, 4βHC and 7KETO) were simultaneously detected by LC-MS in GES-1 and SULT2B1^−/− GES-1 cells with or without adenovirus-mediated SULT2B1b overexpression. b The GES-1 cells were pretreated with 1 μmol/L of 24(R/S),25-EC and 27 HC for 12 h and then with IGF-1 (100 ng/mL) or EGF (100 ng/mL) for an additional 24 h. Cell proliferation was detected by the EdU incorporation assay. EdU fluorescence was normalized to that of Hoechst 33342. c, d GES-1 cells were pretreated with 1 μmol/L of 24(R/S),25-EC and 27HC for 12 h and then with IGF-1 (100 ng/mL, c) or EGF (100 ng/mL, d) for 20 or 90 min. The phosphorylation levels of AKT were detected by Western blotting. The p-AKT/t-AKT ratios are plotted. e GES-1 cells were treated with 1 μmol/L 24(R/S), 25-EC and 27 HC for 12 h. The mRNA levels of the genes involved in gastric epithelial function were measured by qRT-PCR. f CCND1 and CCNA2 protein expression levels were detected by Western blotting. g GES-1 cells were treated with oxysterol (1 μmol/L) and 3-MCA (2 μg/mL) for 10 days. Oxysterol and 3-MCA were added into the medium every 2–3 days. The cell morphology is presented. The mRNA levels of pro- and anticancerous genes in the absence or presence of oxysterol/3-MCA treatment for 10 days were detected by qRT-PCR. * P < 0.05, ** P < 0.01