Figure 3.

Endoplasmic reticulum (ER) and oxidative stress involvement in GTN and z-VAD-fmk-induced MDA-MB-231 necroptosis. ROS production was measured by dihydroethidium (DHE) (A), and 2’,7’-dichlorodihydrofluorescin diacetate (DCFH-DA) (B) represents superoxide anion radicals and hydrogen peroxide radicals, respectively, followed by fluorescent intensity detection. N-acetylcysteine (NAC) is an antioxidant, which was used to inhibit intracellular ROS production. Cytosolic calcium ion level in GTN-treated cells in the presence or absence of z-VAD-fmk, which was stained by Fluo3-AM dye and fluorescence intensity was measured by FACS (C). From three independent experiments in triplicate, the significance of the statistical values compared to the un-treatment group are marked with * p < 0.05; ** p < 0.01, and compared to GTN alone as # p < 0.05 and ## p < 0.01.