DMOG induces intracellular alkalinization associated with increased CAR activity and altered expression of CAR isoforms. (A) Intracellular pH of BMDACs. (B) Extracellular pH of BMDACs. (C) Quantitative RT-PCR analysis of Na⁺/H⁺ exchangers and active CAR isoforms in BMDAC-D. Gray shaded area represents ± 2-fold difference in BMDAC-D versus BMDAC-V. CAR isoforms: (c) indicates cytosolic; (mt), mitochondrial; (mb), membrane-bound; and (s), secreted. (D) CAR activity of BMDAC homogenates was measured using a colorimetric assay and normalized to the total amount of protein. (E) Effect of CAR inhibition on BMDAC viability under low pH (6.4), glucose (0.56mM), and hypoxia (1% O₂). BMDACs were incubated with the CAR inhibitor acetazolamide (1nM to 500μM) for 12 hours; viability was measured with the LIVE/DEAD Violet fluorescent dye and flow cytometry. (A-E) Cells were cultured for 4 days in the presence of vehicle (V) or DMOG (D) before analysis. *P < .05, BMDAC-V vs BMDAC-D, by Student's t test or Bonferroni post hoc comparisons after 1-way ANOVA. Data are mean ± SEM (n = 3).