Figure 7. Cytosolic peroxiredoxins cooperate in controlling mitochondrial H₂O₂ release.

• A, B
  Peroxiredoxin levels in peroxiredoxin 1 (PRDX1 KO) and peroxiredoxin 2 (PRDX2 KO) knockout cells. The upper subpanel depicts the guides used to generated the KO cell lines. Lysates of the indicated cell lines grown in glucose‐containing medium were analyzed by immunoblot (A) and quantitative proteomics (n = 4, technical replicates) (B). The right subpanel of B lists the identified proteins belonging to cellular antioxidative systems which are not altered for PRDX1 KO or PRDX2 KO, respectively. In both quantitative proteomics experiments, 4,403 proteins were detected in total.
• C
  HyPer7 steady state ratios of the indicated cell lines. HyPer7 probes were targeted to the indicated compartments of the respective cell lines. The cells were grown in glucose‐containing medium. The numbers of cells per experiment for each dataset can be found in Appendix Table S6. In the boxplot, the central band is the median. The lower and upper hinges correspond to the first and third quartiles. The upper whisker extends from the hinge to the largest value no further than 1.5 times the inter‐quartile range, whereas the lower whisker extends from the hinge to the smallest value no further than 1.5 times the inter‐quartile range.
• D
  Response of HyPer7 probes targeted to indicated compartments to treatment of mtDAO‐expressing cell lines with 4 mM of D‐Ala (black, wild type; red, PRDX1 KO; blue, PRDX2 KO; cell generated with the piggyBAC system). The cells were grown in glucose‐containing medium. Solid line represents average, points colored in the lighter version of the respective color are the corresponding ratios measured in individual cells.
• E
  Response of HyPer7 probes targeted to indicated compartments to treatment with 25 µM of exogenous H₂O₂. Indicated cell lines (black, wild type; red, PRDX1 KO; blue, PRDX2 KO) were grown in glucose‐containing medium. Solid line represents average, points colored in the lighter version of the respective color are the corresponding ratios measured in individual cells.

Data information: The numbers of cells per experiment for each dataset can be found in Appendix Table S6. As most of the data were not normal distributed, instead of a t‐test, a Wilcoxon/Mann–Whitney‐U‐test was performed and samples were compared in pairs. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.

Source data are available online for this figure.