Fig. S5.

Mitochondrial respiration of isolated mitochondria from synchronized NIH 3T3 throughout the circadian cycle. NIH 3T3 cells were synchronized with a Dexamethasone (Dex) shock as previously described (27) and harvested at 4-h intervals from 24 to 44 h after the Dex shock. (A) Total RNA was prepared, and mRNA expression levels of Bmal1 and Per2 were determined by quantitative real-time PCR. Data are presented as relative expressions, with means ± SEMs of three independent experiments. (B) Mitochondria were isolated from NIH 3T3 cells as described in Materials and Methods. Oxygen consumption rates (OCRs) of isolated mitochondria were quantified using the Seahorse Flux Analyzer in the presence of palmitoyl CoA and carnitine or pyruvate and malate as indicated. OCR measurements are presented in picomoles per minute as means ± SEMs of thee independent experiments.