Figure 1

Global metabolism is increased upon loss of Rev-erbα. (A) Rev-erbα gene expression in Rev-erbα KO and WT MEFs. n.d. denotes not detected. (B) Western blot analysis to measure Rev-erbα protein levels in WT and Rev-erbα KO MEFs. β-actin was used as a loading control. Densitometry analysis of Rev-erbα levels after normalization to β-actin in Rev-erbα KO and WT MEFs. Minimum of n = 5 independent experiments. n.d. denotes not detected. (C) Oxygen consumption rate (OCR) trace from the Seahorse mitochondrial stress test after sequential injection of oligomycin (Oligo), FCCP and antimycin A/rotenone (Anti/Rot). Levels of basal respiration calculated in Rev-erbα KO and WT MEFs. (D) Extracellular acidification rate (ECAR) trace from glycolysis stress tests after sequential injection of glucose (Glu), oligomycin (Oligo) and 2-deoxy-d-glucose (2-DG). Levels of basal glycolysis calculated in WT and Rev-erbα KO cells. (E) Proton efflux rate measured from Seahorse glycolytic rate assays. Levels of basal and compensatory glycolysis were calculated in WT and Rev-erbα KO MEFs. All Seahorse assay traces were normalized to 10,000 cells. Seahorse calculations represented as a ratio to WT group. N = 3–4 independent experiments. Error bars represented as mean ± SEM. *p < 0.05, ***p < 0.001 versus WT.