Figure 7.

25-HC regulates multiple signaling pathways. (A) AGS cells were treated with the indicated concentrations of 25-HC for 24 h or treated with 5 µM 25-HC for the indicated times before total cellular proteins were extracted for western blot analysis to detect the p-Erk1/2, Erk1/2, p-p38, p38, p-SAPK/JNK, SAPK/JNK, p-AKT, AKT, p-PI3K p85/p55, PI3K p85/p55, p-STAT3, STAT3, p-NF-κB p65, NF-κB p65 expression levels. Results were representative of 3 independent experiments. (B) AGS cells were transfected with NF-κB luciferase reporter plasmid before the cells were treated with 25-HC. After 24 h, the luciferase activities were determined by using the Bright-Glo luciferase assay system. Results were obtained from 3 independent experiments and are expressed as the means ± SEM. Statistical significance was determined by one-way ANOVA followed by Dunnett's test. Representative images are presented. ^*P<0.05 and ^***P<0.001. 25-HC, 25-hydroxycholesterol; GC, gastric cancer.