Fig. 7: NO directs metabolic reprogramming in vivo.

a Mice were injected i.p. with thioglycolate and challenged after 3 days with injections of IFNγ or PBS i.p. 12 h later, mice were administered LPS i.p. and euthanized in the subsequent 12 h. Peritoneal lavage fluid was harvested and cells isolated: both processed for metabolic studies. b Nitrite levels were quantified by Griess reaction performed on concentrated lavage fluid in comparison with 2 × 10⁶ LPS/IFNγ stimulated BMDM-derived culture media. Results are shown as total nmol. (n > 3 for each group). c Mice were challenged as in a and either Ly6G + or Cd11B + cells were isolated from peritoneal lavage and subjected to metabolite extraction and subsequent ESI-LC/MS-MS analysis. Normalized peak area of citrulline is shown. d–g Quantified metabolites in Cd11b + Ly6G- isolated from WT mice challenged for Shwartzman reaction (n = 6) (“t” represents mice injected with thioglycolate alone). h Aconitase 2 activity in total CD11b⁺ cells from mice undergoing Shwartzman reaction (n = 3). Data were analyzed by Student’s t-test. i WT and Nos2^−/− mice were challenged as in a and peritoneal lavage was concentrated, extracted and analyzed by ESI-LC/MS-MS analysis. Absolute amounts as ng/total μg protein in lavage fluid are shown. Data were analyzed by two-way ANOVA with Sidak’s post-tests (n = 8). Shown p-values indicate WT vs. Nos2^−/− comparisons in IFNγ + LPS condition. j Quantified metabolites in Cd11b⁺Ly6G⁻ isolated from challenged WT and Nos2^−/− mice. The dotted line represents the average level of metabolite in thioglycolate alone-group. Data were analyzed by Student’s t-test (n = 3). k CI in gel activity and SDS IBs for CI (NDUFS1) and CII (SDHA) in total CD11b⁺ cells from challenged WT vs. Nos2^−/− mice (n = 2). Source data are provided as a Source Data file.