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. 2020 Apr 14;11:1783. doi: 10.1038/s41467-020-15604-2

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© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2020

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Fig. 3 — a NAD⁺ and NADH in log phase wild-type (WT) or ∆gpmA Salmonella grown in aerobic MOPS-glucose media. Mean ± SD; N = 8. b Aerobic respiration of log phase Salmonella grown in MOPS-glucose media was measured by polarography. Mean ± SEM; N = 6 except 4 for the complemented strain. c NADH-I and II dehydrogenase enzymatic activities of log phase Salmonella in MOPS-glucose media. Mean ± SD; N = 12. d Susceptibility of log phase Salmonella in MOPS-glucose media to 400 μM H₂O₂. Mean ± SD; N = 3. e NADH dehydrogenase enzymatic activity was determined in membranes of aerobic Salmonella in LB. Some cells were treated with 50 μM H₂O₂ for 10 min. Mean ± SD; N = 6. f Intracellular pH (pH_i) of log phase Salmonella grown aerobically in MOPS-glucose media, pH 7.2, after treatment with 400 μM H₂O₂ as measured ratiometrically with the pHluorin. When indicated, the ΔpH was collapsed with 40 mM of the protonophore benzoate. Mean ± SD; N = 3 for ΔgpmA, N = 4 for WT. g, Proton motive force as measured spectrophotometrically with the JC-1 probe in stationary phase Salmonella in aerobic MOPS-glucose media. Some cells were pretreated with 400 μM H₂O₂ or 20 μM CCCP for 20 min at 37 °C. N = 3. h Intracellular pH (pH_i) of Salmonella in macrophages derived from C57BL/6 or nox2^−/− mice as measured by fluorescence microscopy. pHluorin⁺ Salmonella were imaged every 2 min p.i for 30 min. Mean ± SEM; N = 5 for B6, N = 7 for nox2^−/−. i. Phagosomal pH in infected macrophages was monitored for 1 h after infection with FITC- and Alexa-fluor647-labeled Salmonella. Mean ± SD N = 3. j Killing of Salmonella by periodate-elicited macrophages from C57BL/6 or nox2^−/− mice 18 h after infection. Mean ± SD; N = 6. k Effects of depolarization on H₂O₂ killing. Some specimens were treated with 400 μM H₂O₂ and/or 20 μM CCCP. Mean ± SD; N = 9 except for CCCP where N = 3. Data were analyzed by unpaired, one-tail t-test (a–c, h), or one-way (d, k) or two-way (e, j) ANOVA. *, **, ***, ****p < 0.05, 0.01, 0.001, 0.0001, respectively. Source data are included in Source Data file.