Figure 3

The activation of GPR30 induced mitochondrial-related apoptosis. (a) SkBr3 and MDA-MB-231 cells were treated with increasing concentrations of G-1 for 48 h, stained with annexin V-FITC and PI, and then analyzed by flow cytometry for cell apoptosis. (b) SkBr3 cells were treated with G-1 as the indicated concentrations for 24 h, and then JC-1, the mitochondria-specific dye, was added to measure the membrane polarity (ΔΨm) and cell apoptosis. Apoptotic cells mainly show green fluorescence (FITC), while healthy cells show red fluorescence (PE). (c) SkBr3 cells were treated with various concentrations of G-1 for 4 h, and then loaded with CM-H₂DCFDA. The fluorescence intensity was measured by FCM. (d) SkBr3 cells were treated with G-1 as the indicated concentrations for 48 h, and then Bcl-2, Bax, Bim, and caspase-3 protein expression levels were analyzed by western blotting. Data were presented as means±S.D. of three independent experiments