Figure 4

Probe uptake and effect of mitochondrial membrane potential. (a) The uptake of DCP-NEt₂C and DCP-Rho1 into live cells was monitored over time by imaging analysis. The change in fluorescence intensity was fit to a bi-exponential equation to extract the kinetic parameters of uptake into cells. (b) The uptake of probes in live and fixed cells measured by flow cytometry was linearly correlated with the probe concentration for both DCP-NEt₂C and DCP-Rho1, the symbols represent the mean fluorescence for each concentration. (c) The effect of pyruvate concentration and FCCP on mitochondrial membrane potential using isolated mitochondria. Compared to 0 mM pyruvate: NS = effect size zero or small; *p < 0.005, effect size medium; **p < 0.005, effect size large. (d) The effect of pyruvate concentration and FCCP on DCP-NEt₂C uptake in isolated mitochondria. Compared to 0 mM pyruvate: NS = effect size small, compared to no probe: ***p < 0.005, effect size very large. (e) The effect of pyruvate concentration and FCCP on DCP-Rho1 uptake in isolated mitochondria. Compared to 0 mM pyruvate: NS = effect size small, *p < 0.005, effect size medium, compared to no probe: ***p < 0.005, effect size very large, (f) The effect of FCCP on mitochondria membrane potential in live cells using flow cytometry. (g) Composite images of the DCP-NEt₂C probe (cyan color) or DCP-Rho1 (magenta) with mitochondrial marker TOMM20 (green) in methanol fixed A549 cells; the corresponding ICA plots from colocalization analyses are shown in Fig. S4.