Fig. 3: Microdomain-specific ROS diffusion to canonical mitochondrial toxins.
From: All-optical spatiotemporal mapping of ROS dynamics across mitochondrial microdomains in situ
a Schematic of mitochondrial ETC toxins producing ROS at distinct sites. b Representative images of dually excited HyPer7 and pseudo-simultaneous imaging of HyPer7 488 and TMRM in response to drug treatments in HEK293T cells. The scalebar denotes 5 µm. c TMRM responses of cells treated with increasing concentrations of rotenone and FCCP. One-way Kruskal-Wallis ANOVA with Dunn’s post-hoc correction, N = 40 – 193 cells, mean ± SD. d Relative HyPer7 ratio responses to ROS produced at complex I through increasing rotenone concentrations. One-way Kruskal-Wallis ANOVA with Dunn’s post-hoc correction, N = 15 – 44 cells, mean ± SD. e TMRM responses of cells treated with increasing concentrations of antimycin A and FCCP. One-way Kruskal–Wallis ANOVA with Dunn’s post-hoc correction, N = 9–114 cells, mean ± SD. f Relative HyPer7 ratio responses to ROS produced at complex III through increasing antimycin A concentrations. Ordinary one-way ANOVA with Dunnett’s post-hoc correction, N = 15–43 cells, mean ± SD. g Equivalents of exogenously added H2O2 needed to reach HyPer7 response to 100 µM antimycin A, normalized to microdomain-specific HyPer7 responses. One-way Kruskal-Wallis ANOVA with Dunn’s post-hoc correction, N = matrix-HyPer7: 13; IMS-HyPer7: 32; OMM-HyPer7: 31 cells, mean ± SD. h Equivalents of exogenously added H2O2 needed to reach HyPer7 responses due to maximal concentrations of rotenone and antimycin A in matrix-HyPer7 expressing cells. Two-tailed, Mann-Whitney test, mean ± SD, N = 31 rotenone, 15 antimycin A treated cells. *p < 0.05, **p < 0.01, ***p < 0.001. Exact N values are provided in the Source Data file.
