Mitochondrial complex activity in permeabilized cardiomyocytes following Thiamet-G (ThG). AC16 cells (10 × 106/well) were plated on 96-well plates and treated with increasing doses of ThG (1, 3, and 5 µM) for 6 h. Cells were placed in MAS buffer for the permeabilization assay. First basal oxygen consumption rate (OCR) was established, and then, 5 mM pyruvate, 2.5 mM malate, and 1 mM ADP were injected; followed by 1 µM rotenone, 10 mM succinate, and 1 mM ADP; and then 10 µM antimycin A. A: indexes of increasing ThG concentration on complex I-dependent activity were calculated. B and C: complex II-dependent activity was determined (B) and complex IV dependent activity (C). Data are means ± SE; n = 3–5 replicates per sample. *P < 0.05, different from untreated control. #P < 0.05, different from 1 µM ThG treatment. ^P < 0.05, different from 3 µM ThG treatment.