Extended Data Fig. 2: Imaging redox-dependent changes in mitochondrial cysteine oxidation.
a, Amide derivative of Wittig reagents exists predominantly in protonated form, setting stage for enrichment and detecting S-sulfenylation in mitochondria. b, Structure of the mitochondrial targeting sulfenic acid probe WYneN10 with enhanced lipophilicity. c, Live HeLa cells were incubated with BDP-WYneN10 (500 nM) and MitoTrackerTM Deep Red FM (100 nM) in DPBS. After 10 min, confocal images were taken. A scale bar of 20 µm is shown. R, Pearson’s correlation coefficient. d, BDP-WYneN10 tagged S-sulfenylated proteins with fluorescence inside mitochondria. e, BDP-WYneN10 fluorescence responded to external oxidative stress (0-5 mM H2O2) in live A549 cells (n = 4 areas from one representative experiment). f, WYneN10 disrupted mitochondrial respiration in A549 cells to a greater extent than other WYne probes (50 µM) (n = 12 biological replicates). OCR, oxygen consumption rate. Data in e-f are presented as box plots (maximum, 75%, median, 25%, minimum). P values were calculated using a two-tailed t-test. ns, not significant, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001.
