Fig. 2.
RGS11 prevents doxorubicin-induced oxidative stress, mitochondrial dysfunction, and cell death in human cardiomyocytes. (A) Human AC-16 cardiomyocytes were treated with doxorubicin (3 μM, 16 h), 5-FU (500 μM, 16 h), or oxaliplatin (0.06 mM, 16 h) and immunoblotting performed probing for RSG11 (n = 3). (B–H) Control (scramble plasmid) or RGS11 OE AC-16 cells were treated with doxorubicin (3 μM, 16 h) ± pre-treatment with mitochondrial calcium uniporter inhibitor Ru360 (50 μM, 1 h) or mitochondrial permeability transition pore (mPTP) blocker cyclosporin A (0.2 mM, 45 min) where indicated. (B) Immunoblotting for RGS11 (n = 6).(C) GPX activity (n = 5) and SOD activity (n = 5). (D) Mitochondrial Ca2+ flux (n = 5). (E) Mitochondrial membrane potential (ΔψM; n = 5). (F) Caspase-3 cleavage (n = 5). (G) Apoptosis (cytoplasmic histone-associated DNA fragments; n = 5). (H) Annexin V+ cells (n = 5). β-Actin serves as a loading control for western blots. Immunoblots are accompanied by a densitometric quantification wherein expression is normalized to the corresponding control group. Data were analyzed by one-way ANOVA with Sidak's post-hoc test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ns = not significant. Data are presented as mean ± SEM.