Fig. 2

Enhanced proliferation inhibition by GPR119 agonists in breast cancer cells. a and b Synergistic effects of GPR119 agonist on cell proliferation inhibition by gefitinib in MCF-7 a and MDA-MB-231 cells b. Isobole model was used to evaluate drug combination effect. c Apoptosis induction by gefitinib with MBX-2982. Apoptosis was determined by PI and annexin V staining. The stained cells were analyzed by flow cytometry in MCF-7 cells. Annexin V-positive pro-apoptotic and late-apoptotic cells were counted by BD CellQuest Pro software. d Caspase3/7 activity. MCF-7 cells were preincubated with DEVD-NucViewTM 488, and exposed to gefitinib and MBX-2982 for 60 h, and caspase-3/7-selective green fluorescence was monitored. Green fluorescence intensity was calculated by Incucyte® ZOOM basic analyzer. e PARP cleavage by gefitinib with MBX-2982. MCF-7 cells were incubated in the presence of gefitinib or gefitinib with MBX-2982 for 48 h. Western blot analyses were performed to determine PARP cleavage. f Expression of Bcl-2 and Bax. g Expression of cleaved caspase-8 (active form). MCF-7 cells were incubated with 10 μM gefitinib, 10 μM MBX or gefitinib/MBX for 24 h. Cell cycle analysis h and cell cycle-associated protein expression i. MCF-7 cells were treated with MBX-2982 (MBX) for 24 h and fixed with ethanol. Cell cycle was analyzed by propidium iodide (PI) staining in MCF-7 cells. For the quantification of protein expression of p53, p21 and p27, MCF-7 cells were incubated with 1-10 μM MBX-2982 for 24 h. Data represent the mean ± S.D. (n=3). ** p < 0.01, *** p < 0.005 significant difference versus control group