Fig. 3: The effects of Bach1 expression on pulmonary inflammation is reflected in the modulation of the cell-intrinsic oxidative response.

a–i, Bach1^−/− and WT mice were infected at high-dose Mtb and euthanized at 28 dpi. a, tSNE analysis of myeloid cells in the lungs of Mtb-infected mice (n = 5 per group). b, Frequencies and numbers of AM (Live/CD45⁺/DUMP^-/Ly6G⁻/CD45iv^neg/CD88⁺CD64⁺/CD11b^−/low/CD11c⁺/Siglec-F⁺), IM (Live/CD45⁺/DUMP⁻/Ly6G⁻/CD45iv^neg/CD88⁺/CD64⁺/CD11b^hi/CD11c^-/low/Siglec-F⁻) (two-tailed, Mann–Whitney U-test), c, total neutrophils (Live/CD45⁺/DUMP⁻/CD11b⁺Ly6G⁺) (two-tailed Mann–Whitney U-test) and d, parenchymal neutrophils (Live/CD45⁺/DUMP⁻/CD11b⁺Ly6G⁺/CD45iv⁻) (two-tailed Mann–Whitney U-test) were enumerated; DUMP: TCRβ, TCRγδ, NK1.1, B220. e, Lipid peroxidation (LAA staining) (two-tailed Mann–Whitney U-test) and f, Gpx4 levels (over the average MFI of the WT) (two-tailed Mann–Whitney U-test) in live IM and neutrophils. b–f, Pooled data from 3 independent experiments are shown (WT n = 13 vs Bach1^−/− n = 12). g, Total antioxidant status (n = 16 per group, pooled from 3 experiments; two-tailed Mann–Whitney U-test). h, Glutathione measured in lung homogenates (WT n = 20 vs Bach1^−/− n = 22, pooled from 4 experiments; two-tailed Mann–Whitney U-test). i, Heatmap visualization of 10 cytokines/chemokines measured in lung homogenates. Pooled data from 3 independent experiments are shown (WT n = 16 vs Bach1^−/− n = 14). j, Schematic of mixed BM chimaeric protocol. Mice were euthanized at 13 dpi. k, Analysis of cell frequency (two-tailed Wilcoxon-matched pairs), l, lipid peroxidation (two-tailed Wilcoxon-matched pairs) and m, Gpx4 expression (two-tailed Wilcoxon-matched pairs) in AM, IM and neutrophil populations in the lungs of Mtb-infected mixed (WT/Bach1^−/−) BM chimaeras. Results shown are pooled from 2 independent experiments (n = 15 per group; Wilcoxon-matched pairs). Data shown in b–m are presented as mean ± s.e.m. and each symbol represents an individual animal.

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