Fig 2: STAT1-dependent control of Cybb expression and CYBB protein levels in neutrophils.(A) Heatmap of Cybb, Cyba, Ncf1, Ncf3, Ncf4, Nox1, Nox3, and Nox4 expression in neutrophils sorted from Stat1−/− (purple symbols) and Stat1+/+ (black symbols) mixed bone marrow chimeric mice (n = 6). Genes with a STAR* are DEG. Each lane is an independent biological replicate. (B) qRT-PCR of Cybb mRNA expression in neutrophils sorted from Stat1−/− (purple symbols) and Stat1+/+ (black symbols) mice. (C) ATAC-seq analysis of neutrophils were sorted from Stat1−/− (purple peaks) and Stat+/+ (black peaks) mice. Gene tracks show chromatin accessibility at the Cybb and Gbp2 locus. (D) Bone marrow and lung neutrophils were sorted from Stat1−/− (purple peaks) and Stat1+/+ (black peaks) mice and processed for CUT&RUN. Gene tracks show STAT1 signal as normalized fragment pileup (y-axis) plotted by genome position (x-axis). The shaded box highlights a putative STAT1 binding site at the Gbp2 promoter region. Gene tracks show H3K4me3 ChIP-seq signal as normalized fragment pileup (top rows; green). (E - H) Western blot and quantitation of (F) CYBB (G) CYBA (H) NCF4 vs GAPDH protein levels in lung neutrophils sorted from Stat1−/− (purple symbols) and Stat1+/+ (black symbols) mice neutrophils from 4-5 mice were pooled to obtain sufficient protein for Western blot analysis in each biological replicate. (A - H) Infection dose: 3 × 107 CEA10 conidia via intratracheal route analysis 72 h pi. (C) Data were calculated by Kruskal-Wallis nonparametric test for multiple comparisons for each group compared with control group. (G – I) Data are representative of 2 experiments. Dots represent individual mice. Statistical analysis: Mann-Whitney test. See also Figure S4.

Fig 3: Expression of p22phox, a subunit of NADPH oxidase, in kidney tissues of Ob-NAFLD rats. Rat kidney sections from lean control (LC), lean treated with metformin (L + M), obesity associated NAFLD (Ob-NAFLD), and metformin-treated Ob-NAFLD (Ob-NAFLD = M) groups were employed for immunohistochemistry of p22phox, a subunit of NAPDH complex and a marker of reactive oxygen species production. (A) Representative immunohistochemistry images showing p22phox+ cells in rat kidneys of the following: (a) LC; (b) L + M; (c) Ob-NAFLD; and (d) Ob-NAFLD + M groups. Scale bars = 100 μm. (B) Graph showing quantification of p22phox+ cells within 10 microscopic fields (40×) across experimental groups. Data are expressed as mean ± SEM (n = 5 per group). Statistical significance between the two groups was determined by a non-parametric Mann–Whitney U test (** p < 0.01).