REV-ERBβ knock down drives mitochondrial gene expression <i>in vitro</i>.

<p><b>(A)</b> Mitochondrial biogenesis determined by flow cytometry (FACS) using MitoTracker Red staining in empty vector and REV-ERBβ shRNA overexpressing C2C12s. FACS panels (left) indicate percent of cells retrovirally transduced. FACS panels (middle) demonstrate knock down of REV-ERBβ enhances MitoTracker Red staining. Graph (right) depicts median fluorescent intensity (MFI). <b>(B)</b> qRT-PCR analysis of genes demonstrating cells had terminally differentiated. <b>(C)</b> Mitochondrial DNA (mtDNA) content demonstrated by the ratio of mtDNA to nuclear DNA (nucDNA) in CD8 shRNA versus REV-ERBβ shRNA overexpressing C2C12s. nucDNA was measured using ß-actin (<i>Actb</i>) primers. <b>(D)</b> qRT-PCR analysis demonstrating the knock down of REV-ERBβ and its effects on RORα, REV-ERBα, and genes encoding proteins involved in fatty acid oxidation and cytokine production. <i>18s</i> was used as the internal control. (n = 3 per condition). Values are mean±s.e.m. Data representative of 4 independent (Panels A, B, D) or two independent (Panel C) experiments demonstrating similar results. Statistical significance was assessed using Student’s two-tailed <i>t-</i>tests. <i>*p</i><0.05, ** <i>p</i><0.01, *** <i>p</i><0.005, **** <i>p</i><0.001.