Fig. 4: Transient mtDNA depletion induced by mitoARCUS3.2 does not impact respiration in high-percentage m.3243G mutant cybrid cells grown in glucose or galactose.
a–l, Cell lines C (95% mutant) and B (85% mutant) were nucleofected with mRNA encoding either MTS-GFP or mitoARCUS3.2 at a dose of 1 × 105 mRNA copies per cell. Cellular DNA was collected at day 1 for mtDNA heteroplasmy and mtDNA copy number analysis, and live cells were analysed for respiration. mtDNA heteroplasmy, normalized to mtDNA copy number of control in cell line C (95% mutant) cultured in glucose-containing medium (a). Seahorse Cell Mito Stress Test for cells described in a (b). Basal respiration for cells described in a, normalized to control (c). Maximal respiration for cells described in a, normalized to control (d). mtDNA heteroplasmy, normalized to mtDNA copy number of control in cell line B (85% mutant) cultured in glucose-containing medium (e). Seahorse Cell Mito Stress Test for cells described in e (f). Basal respiration for cells described in e, normalized to control (g). Maximal respiration for cells described in e, normalized to control (h). mtDNA heteroplasmy, normalized to mtDNA copy number of control in cell line C (95% mutant) cultured in galactose-containing medium (i). Seahorse Cell Mito Stress Test for cells described in i (j). Basal respiration for cells described in i, normalized to control (k). Maximal respiration for cells described in i, normalized to control (l). Data encompass three independent experiments and are shown as mean ± s.d. Statistical analysis was performed using a two-tailed t-test on raw data. NS, P > 0.05; all other P values indicated.
