Figure 4.

(a) Differences in protein expression of Bcl-2, Bax, and cleaved-caspase 3 after treatment with 10 mM Ac or 5 mM Bu in GES-1 cells. (b-c) Differences in protein expression of Cyclin D1, Bcl-2, Bax, and cleaved-caspase 3 after treatment with 10 mM Ac or 5 mM Bu in GPR109A-knockdown GC cells. (d-g) Changes in cell proliferation (d-e) and apoptosis (f-g) were observed in MGC-803 and HGC-27 cells after siRNA-mediated knockdown of GPR109A and treatment with Ac or Bu. (h) Co-immunoprecipitation confirmed the interaction between GPR109A and HOPX proteins. (i) Differences in HOPX protein expression were detected by IHC in cancer and adjacent tissues of GC patients. (j) Quantification of the results of (I). (k-l) Differences in the co-localization of GPR109A and HOPX between the GC cell lines MGC-803 and HGC-27 after siRNA-mediated knockdown of GPR109A combinated with 5 mM Bu. (m) Differences in the expression of apoptosis-related proteins Bcl-2, Bax, and cleaved-caspase 3 were observed in MGC-803 and HGC-27 cells after siRNA-mediated knockdown of GPR109A and HOPX, knockdown of GPR109A, or overexpression of HOPX. (n) Differences in the expression of apoptotic and anti-apoptotic-related proteins were scrutinized after (M) combinated with 5 mM Bu. (o-p) Changes in cell apoptosis rates after the (M) treatment. (q-r) Changes in cell apoptosis rates after the (N) treatment. Data indicate the mean ± SD. *p < .05, **p < .01, and ***p < .001, by 2-tailed Student’s t test or one-way ANOVA.