DMOG elicits transcriptional and metabolic responses in BMDACs. (A) BMDACs were cultured in the presence of vehicle (V) or DMOG (D). Mitochondrial mass (top panels), ROS (middle panels), and glucose uptake (bottom panels) were measured in BMDACs by flow cytometry after staining with the fluorescent dyes nonyl acridine orange (NAO), 2′,7′-dichlorodihydrofluorescein diacetate (H₂DCFDA), and 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG), respectively (n = 4-6). (B) Mitochondrial DNA content relative to nuclear genomic DNA was determined by quantitative PCR (n = 6). (C) Oxygen consumption (JO₂) was analyzed in BMDACs suspended in serum-free EGM2-MV media (n = 6). (D) Quantitative RT-PCR analysis of HIF-1 target genes encoding metabolic regulators in BMDACs cultured in vehicle or DMOG for 4 days (n = 4-6). ETFA and TBP are not HIF-1 targets and were used as negative controls. Gray shaded area indicates ± 2-fold of control. (Inset) Immunoblot assay showing BNIP3, LDHA, and PDK1 protein levels. (E) Extracellular lactate content of BMDAC cultures (n = 9). (F) HIF-1α protein expression in BMDACs was determined by immunofluorescence using flow cytometric analysis (left panel). Gray histogram corresponds to the negative control omitting the primary antibody. Data are mean ± SEM (n = 6; right panel). *P < .05, DMOG vs vehicle, by Student t test in all panels of this figure. Data are mean ± SEM.