Figure 4

ρ0 and ρ + mutant recipient cells have restored respiration with transferred CAP-R mitochondria. (a–c) Seahorse Extracellular Flux analysis and quantification of ATP levels contributed by mitochondria (ATPmito) and glycolysis (ATPglyco). Cells were cultured in uridine-deficient, CAP-supplemented media. (a) Analysis of CAP-R 501-1 mitochondrial donor, L929 ρ0 recipient, and L929 ρ0 + CAP-R 501-1 cells. (b) Analysis of CAP-R 501-1 mitochondrial donor, Δmt-ND4, and Δmt-ND4 + CAP-R 501-1 bulk culture cells from three independent transfers. (c) Analysis of Δmt-ND6, CAP-R 501-1, and Δmt-ND6 + CAP-R 501-1 bulk cultures from two independent transfers. (a–c) Two-tailed, unpaired Student’s t-test comparing samples to L929 ρ0, Δmt-ND4, or Δmt-ND6. * represents significance for ATPmito and ‡ represents significance for ATPglyco. * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. ‡ < 0.05, ‡ ‡ < 0.01, ‡ ‡ ‡ < 0.001, ‡ ‡ ‡ ‡ < 0.0001. The bar height denotes average of 4 replicates and the error bars are the standard deviation. (d) Seahorse Extracellular Flux analysis and quantification of ATPmito and ATPglyco in L929 ρ0, Δmt-ND4, CAP-R 501-1, L929 ρ0 + CAP-R 501-1 bulk cultures from three independent transfers, and Δmt-ND4 + CAP-R 501-1 bulk cultures from three independent transfers. Cells were cultured in uridine-supplemented, CAP-supplemented media. Two-tailed, unpaired Student’s t-test comparing samples to Δmt-ND4. * represents significance for ATPmito and ‡ represents significance for ATPglyco. * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. ‡ < 0.05, ‡ ‡ < 0.01, ‡ ‡ ‡ < 0.001, ‡ ‡ ‡ ‡ < 0.0001. There were no statistically significant differences when comparing the samples to L929 ρ0 cells. The bar height denotes average of 5 replicates and the error bars are the standard deviation. (e) Schematic showing summary of ρ + mitochondrial transfer efficiency in a given selection condition.