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Fig. 2 | BMC Cancer

Fig. 2

From: Increased sensitivity of African American triple negative breast cancer cells to nitric oxide-induced mitochondria-mediated apoptosis

Fig. 2

Nitric oxide induced mitochondria mediated apoptosis in AA breast cancer cells. a-b) Cells were seeded to confluence and treated with DETA NONOate (0.5-1mM) for 48h. Immunoblot analysis was performed for cleaved caspase-3 (19 and 17kDa band), total Bax (Intact Bax 20kD and cleaved Bax 18kD band) and Bcl2. Expression of β-actin was used as housekeeping control. Neither caspase-3 nor Bax cleavage was found in CA cell lines. c) Cells were exposed with DETA-NONOate (1mM) with or without pretreatment (1h) with various concentrations (10-30μM) of Calpain III (Ci III) inhibitor. Data is presented as mean ± SD with significant values presented as ***p < 0.001. d, e) Cells treated with DETA-NONOate and Calpain inhibitor III were subjected to Immunoblot analysis for cleaved caspase-3 and Bax. f) Cells treated with DETA NONOate for 24h (blue curve) or 48h (red curve) were harvested and incubated with Mito Tracker dye to measure mitochondrial membrane depolarization using flow cytometer. Uncoupling agent carbonyl cyanide-4-(trifloromethoxy) phenyl hydrazine (FCCP) was used as positive control (green curve) while untreated cells served as control (black curve)

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