Extended Data Fig. 5: Exposure to hypoxia does not affect the frequencies of circulating neutrophils upon infection, progenitor abundance or N-terminal H2B clipping in mice.
a, Circulating neutrophil abundance obtained by flow cytometry in mice exposed to 10 % O2 for a week followed by a 5-week reoxygenation period prior to subcutaneous infection with S. aureus SH1000 (Hpx + ReNmx). Normoxia counterparts (Nmx) were used as experimental controls. Data expressed as a percentage from CD45+ cells 48 h post-challenge (n = 4 Nmx and n = 5 Hpx + ReNmx). b, Granulocyte monocyte progenitor (GMP) and Common myeloid progenitor (CMP) abundance quantification by flow cytometry in the bone marrow of mice nebulized (HpxLPS + ReNmx) with LPS or not (Hpx + ReNmx) and exposed to 10 % O2 for a week followed by a 5-week reoxygenation period. Normoxia counterparts (Nmx) were used as experimental controls (n = 3 for all experimental conditions). c–e, Quantification of bone marrow proNeu1, proNeu2 and preNeu from mice subjected to 10 % O2 for a week followed by a 3-month reoxygenation period (Hpx + ReNmx) compared to normoxia control mice (Nmx) obtained by flow cytometry as a proportion of Alive/Lin- cells (n = 4 Nmx and n = 3 Hpx + ReNmx). f, g, Mice were nebulized (HpxLPS) or not (Hpx) with LPS and subjected to 10 % O2 for a week, with normoxia counterparts (Nmx) used as experimental controls. Bone marrow preNeu were harvested and lysed followed by chymotrypsin digestion. Liquid chromatography-mass spectrometry (LC–MS) was used to identify histone peptides. f, Total intensity for the peptides corresponding to histone 2 (H2). g, Abundance of N-terminal histone H2B peptide corresponding to amino acids 5-20 in the protein sequence (n = 4 for all experimental conditions). Data as mean ± s.d. Each value represents a mouse, except (b), with 4 mice pooled to obtain each data point. Significant p values depicted (for p < 0.05) and obtained by Shapiro-Wilk normality test followed by two-tailed t-test (a, c, e), 1-way ANOVA (b), two-tailed Mann–Whitney test (d), or 2-way ANOVA (f, g).
