Fig. 3: mitoARCUS3.2 shifts heteroplasmy in 95% of m.3243G mutant cybrid cells, resulting in increased steady-state levels of mitochondrial proteins and increased respiration at day 3. | Nature Metabolism

Fig. 3: mitoARCUS3.2 shifts heteroplasmy in 95% of m.3243G mutant cybrid cells, resulting in increased steady-state levels of mitochondrial proteins and increased respiration at day 3.

From: Efficient elimination of MELAS-associated m.3243G mutant mitochondrial DNA by an engineered mitoARCUS nuclease

Fig. 3

a–e, Cell line C (95% mutant) was nucleofected with either an MTS-GFP control or mitoARCUS3.2 mRNA at tenfold dilutions, starting at 1 × 105 mRNA copies per cell. Cellular DNA was collected at day 3 for mtDNA heteroplasmy and mtDNA copy number analysis (a), protein lysates were collected for immunoblot (b) and live cells were analysed for respiration (c–e). mtDNA heteroplasmy, normalized to mtDNA copy number of control (a). Immunoblots showing MT-CO1, NDUFB8 and alpha-tubulin steady-state levels (b). Seahorse Cell Mito Stress Test (c). Basal respiration, normalized to control (d). Maximal respiration, normalized to control (e). Data encompass three independent experiments and are shown as mean ± s.d. Statistical analysis was performed using one-way ANOVA on raw data. NS, P > 0.05; all other P values indicated.

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