Fig 2: Magnolol increased M2c-like macrophage subpopulations. Changes in immune cell subtypes by magnolol treatment were determined using flow cytometry in splenocytes from either control (con), magnolol (mag), cisplatin (cis), or cis+mag mice. (A) Gating strategy for analysis of macrophages and T cells subsets. Red dots display isotype controls and black dots show the stained cells with specific antibodies. (B) Representative dot plots of CD11b+F4/80+ macrophages, (C) CD8+ and CD4+ T cells. (D) Bar graphs showing mean% of CD11b+F4/80+ macrophages, (E) CD8+ T cells, and (F) CD4+ T cells in CD45+ total leukocytes in splenocytes. (G,H) Analysis of macrophage subtypes. (G) Representative dot plots identifying macrophages as CD163-CD206- M1, CD163-CD206+ M2a, and CD163+CD206+ M2c gated on CD45+CD11b+F4/80+ cells. (H) Bar graphs showing mean% of M1, M2a, and M2c macrophages in CD11b+F4/80+ splenocytes. All data are presented as the mean ± SEM (n = 5). *P < 0.05; **P < 0.01; ***P < 0.001 vs. con, ##P < 0.01; ###P < 0.001 vs. mag, and $$P < 0.01 vs. cis based on the one-way ANOVA Tukey's test. The letters for no significance were not shown.

Fig 3: HIF-1a in NKp46+ cells favors an ILC1 phenotype and controls homeostasis in the SI. (A) The frequency and absolute number (Nb) of small intestinal NKp46+ ILCs together with NK cells in CD45+ population, from HIF-1a KO and HIF-1a WT mice (pooled data of three independent experiments, n = 7); data are mean values ± SEM; two-tailed Student’s t test. (B) Gating strategy, upper panel: ILC characterization, the analysis of alive CD45+ Lin- CD127+ ILCs; ILC1 (NKp46+ ROR?t- T-bet+ NK1.1+ CD49a+); NKp46+ ILC3 (NKp46+ ROR?t+ CD49a-). Lower panel: lymphoid and myeloid cell characterization: the analysis of alive CD45+ cells; neutrophils (CD11b+ Ly6G+); eosinophils (CD11b+ Siglec-F+); macrophages (CD11b+ Ly6C+ F4/80+); dendritic cells (MHCII+ CD11c+); B cells (CD19+ B220+); CD4 cells (TCRß+ CD4+); and CD8 cells (TCRß+ CD8+). FSC, forward scatter; SSC, side scatter. (C) Flow cytometric analysis of alive CD45+ cells, neutrophils (NPs), macrophages (M?), eosinophils (Eos), B cells, total T cells, CD4 cells, CD8 cells, and dendritic cells from steady-state small intestinal tissues from HIF-1a KO and HIF-1a WT mice (pooled data of two independent experiments, n = 7); data are mean values ± SEM; *, P < 0.05 by two-tailed Student’s t test. (D) Flow cytometric analysis of CD4+ cells stimulated with PMA/ionomycin and stained for intracellular IFN-? and IL-22 from HIF-1a KO and HIF-1a WT mice (n = 6); data are mean values ± SEM; two-tailed Student’s t test.

Fig 4: A GATA Switch May Underlie Changes in Gene Expression in the Development of Splenic Red Pulp Macrophages (RPMs)(A) Identification of GATA motifs in accessible chromatin of splenic monocytes (CD11bhi F4/80-), pre-RPMs (CD11bhi F4/80lo), and RPMs (CD11blo/- F4/80hi), within 100 kb of the start sites of genes differentially expressed between WT monocytes and WT pre-RPMs, or WT pre-RPMs and WT RPMs (both log2 fold change > 1, adjusted p value < 0.05). Motif enrichment was performed on accessible chromatin from each splenocyte population from WT and Il1rl1-/- mice and also from accessible chromatin from each splenocyte population that was unique to WT or unique to Il1rl1-/- mice; n = 4 separate mice per group.(B) Relative Gata gene expression (normalized counts, cpm) in splenic monocytes (CD11bhi F4/80-), pre-RPMs (CD11bhi F4/80lo), and RPMs (CD11blo/- F4/80hi) of WT (green) and Il1rl1-/- mice (red); n = 4 separate mice per group. Data are represented as mean ± SEM. **** adjusted p (WT v -/-) < 0.001.(C) Heatmaps showing log2 fold changes of genes differentially expressed in WT and Il1rl1-/- pre-RPMs (adjusted p value < 0.05) between splenic monocytes (CD11bhi F4/80-) and pre-RPMs (CD11bhi F4/80lo), or pre-RPMs (CD11bhi F4/80lo) and RPMs (CD11blo/- F4/80hi), from WT and Il1rl1-/- mice; n = 4 per group.Please see also Figure S7.

Fig 5: The loss of DGAT1 function decreases TG accumulation, LDs, and inflammatory function.a Effect of DGAT1 inhibitor T863 (DGAT1i) on total TG, and b distinct TG species (n = 4 biologically independent samples) in resting (M0) and macrophages stimulated with IFN? + LPS. c Bodipy 493/503 fluorescence visualized by fluorescence microscopy (scale bars represent 10 µm) or d measured by flow cytometry (MFI, median fluorescence intensity) (n = 3 biologically independent samples). e C16 Bodipy uptake (n = 4 biologically independent samples). f Effect of DGAT1i on DG levels in M0 or macrophages stimulated with IFN? + LPS (n = 4 biologically independent samples). g Effect of Dgat1-shRNA (shDgat1) or empty vector (EV) on TG species. h Bodipy 493/503 fluorescence in macrophages transduced with shDgat1 or EV and treated with IFN? + LPS (n = 3 biologically independent samples). i Pro IL-1ß positive macrophages stimulated with IFN? + LPS, plus/minus DGATi or j shDgat1 (n = 4 biologically independent samples). k, l Effect of the loss of DGAT1 function on IL-6 production (k n = 8 and l n = 5 biologically independent samples). m Effect of DGAT1i on phagocytosis of S. aureus (PE-labelled) (n = 4 biologically independent samples). n C57BL/6 mice were injected with DGAT1i, or carrier, 30 min before receiving 8 mg kg-1 of i.p. LPS or PBS. Drop in body temperature was calculated in relation to the initial body temperature. o Bodipy 493/503 fluorescence in large (F4/80+TIM4+), and p small (F4/80+TIM4neg), peritoneal macrophages from the mice mice shown in (n), recovered at 10 h post injection of LPS or PBS (n = 5 biologically independent samples). q Serum IL-1ß at 2 h post injection (n = 4 biologically independent samples). r Serum IL-6 at 10 h post injection (n = 5 biologically independent samples). For a–g and k–m, assays were at 18 h post activation; i, j at 6 h. Data are represented as mean values ± s.e.m. One-way ANOVA with Bonferroni’s multiple comparison test (a, e, f, n); Unpaired two-tailed Student’s t test (d, h–m, p–r). (a, b, e–h, n–r) representative of two or (c, d, i–m) three experiments. Source data are provided as a Source Data file.