Trehalose inhibits hepatocyte respiration and glycolysis to induce the fasting response. (A) Seahorse (XF96) analysis by mito stress test kit assay. Shown is the oxygen consumption rate (OCR) over time (n = 6–8 cell cultures per group), with injection of oligomycin (‘oligo’), FCCP, and rotenone and antimycin (‘Rot/Ant’) at the indicated time points. (B) Quantification of basal OCR in cells treated with or without trehalose (24–72 h). (C) Quantification of maximal (FCCP-stimulated) respiration in cells treated with or without trehalose. (D) Quantification of oxygen consumption that is devoted to ATP production, as determined in hepatocytes after oligomycin treatment. (E) Extracellular acidification rate (ECAR) quantified in cultures analyzed in (A-D) over time. (F) Quantification of basal ECAR, measured immediately prior to oligomycin injection. * P < 0.05, ** P < 0.01, *** P < 0.001 by Student’s T-test with Bonferroni-Dunn post-hoc correction for multiple tests.