Fig. 6.

Retinoic acid-related orphan receptor-α (RORα) promotes mitochondrial phosphorylated (p-)STAT3 Ser⁷²⁷ translocation and mitochondrial function in vitro. A: neonatal rat ventricular myocytes (NRVMs) were infected with lentivirus containing scrambled shRNA (shRNA) or shRNA against RORα (shRORα) and then treated with angiotensin II (ANG II; 200 nM for 24 h). Cell lysates were fractionated and immunoblotted (n = 3 independent experiments). B: NRVMs were infected with doxycycline (Dox)-inducible RORα lentivirus and then treated with ANG II (200 nM) or vehicle for 24 h in the absence (−) or presence (+) of Dox. Representative immunoblot analysis and summary quantitative densitometry (ImageJ) are shown (n = 3 independent experiments). C: after infection and treatment, NRVM genomic DNA was used in quantitative PCR for mitochondrial genes normalized to the nuclear reference gene TATA box-binding protein (Tbp). D: NRVM mitochondria were labeled with MitoTracker red CMXRos. E: H9c2 rat ventricular myoblasts were infected with inducible RORα lentivirus and treated with ANG II (200 nM for 24 h) in the presence or absence of Dox and then stained with dichlorofluorescin (DCF) to assay oxidative stress. F: NRVMs were infected with inducible RORα lentivirus or RORα shRNA. Intracellular ROS was measured using a fluorometric assay. Four-group comparisons used two-way ANOVA; two-group comparisons used an unpaired t-test. *P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001.