Extended Data Fig. 5: Infiltrated neutrophils are the major source of Interleukin-1β and limit excessive energy loss upon β3-adrenergic activation.
From: Neutrophils preserve energy storage in sympathetically activated adipocytes
a. Representative flow cytometry plots (CD45 + CD11b+Thy1.2-CD19- cells) and quantification of each cells in spleen from diphtheria toxin (DT, 20 µg kg−1) treated Neu-WT and Neu-DTR mice 12 h after CL injection (0.5 mg kg−1). (Neu-WT: n = 8; Neu-DTR: n = 10). Quantification of cells in bone marrow 24 h after DT injection. (n = 3 per group). P values Neu-WT vs Neu-DTR. b. Serum NEFA and IL-6 in DT-treated mice after CL injection. (glycerol, 0 h: n = 3,4; 2 h: n = 6; 12 h: n = 10,7; 24 h: n = 6,10. IL-6, 0 h:n = 3,2; 2 h: n = 4,3; 12 h: n = 9,7; 24 h: n = 6,9). P values Neu-WT vs Neu-DTR. c. Inflammatory genes in eWAT (Adgre1) or iWAT (IL-1β, IL-6) from DT-treated mice 12 h after CL injection. (CL 0 h: n = 3,4; CL 12 h: n = 7,8. normalized to Neu-WT 0 h). P values Neu-WT vs Neu-DTR. d. IL-1β levels in supernatants from WT PPDIVs treated with CL-316,243 (1 µM, 12 h) or LPS (0.5 µg ml−1, 12 h), followed by ATP (5 mM, 1 h). (n = 2 per group). e-f. (e) Body weight and (f) food intake from DT-treated mice at indicated time points or 6 h after final CL injection. (CLx2: 2 days; CLx7: 7 days). Food intake was measured for 24 h before and after CL injection. (f, n = 6 per group). P values indicate comparisons between Neu-WT and Neu-DTR. g. Tissue weight of eWAT 6 h after second CL injection with or without DT. (n = 3,4 per group). P values Neu-WT vs Neu-DTR. h. IL-1β levels in eWAT from WT mice after CL injection for the indicated times (CL x2: 2 days injection). (0 h, 6 h: n = 4; 12 h, 24 h: n = 5; x2: n = 6). P values vs 0 h. i. IL-1β levels in eWAT from DT-treated mice 1 h after second CL injection. (CL 0 h: n = 2; CL x2: n = 4). P values Neu-WT vs Neu-DTR or relative vs 0 h. Data are mean ± s.e.m. Each dot (n) represents one biological replicate. Data are merged from three (a, b, c, h) or representative of two (d, f, g, i) independent experiments. Statistics: two-way ANOVA with Sidak’s post hoc test (a (bone marrow), b, c, e, f, g, i); one-way ANOVA with Tukey’s (h); unpaired two-sided Welch’s t-test (a).
