Fig. 6: Loss of Foxp3⁺Tregs enhances diet-induced hypothalamic immune activation.

Tregs were depleted using i.p. administration of diphtheria toxin (DTX) in Foxp3 DTR mice. a Representative FACS plots for macrophages and microglia after maintenance on the HFHS diet for 1 week with or without Treg depletion using Foxp3 DTR mice and ablation with DTX. b Quantification of CD45^hiCD11b⁺ infiltrating macrophages. Data are depicted as box-and-whisker plots (min-to-max values with single data points). N = 6 biological replicates. Two-tailed unpaired Student’s t test with P = 0.0040. c–e Immune activation of CD45^intCD11b⁺ microglia as seen from CD80⁺, CD86⁺ and MHCII⁺ expressing cells. Data are depicted as box-and-whisker plots (min-to-max values with single data points). N = 6−8 biological replicates. f–j Representative FACS plots and quantification of immune activation in infiltrating macrophages as seen from CD80⁺, CD86⁺ and MHCII⁺ expressing cells. Data are depicted as box-and-whisker plots (min-to-max values with single data points). N = 6 biological replicates. k Volcano plot of proteome analysis of FACS-sorted CD45^intCD11b⁺ microglia from Foxp3 DTR mice without (n = 4) or with (n = 6) DTX-mediated Treg depletion exposed to 2 weeks HFHS diet. l, m Activation status and proliferative capacity of brain-residing CD4⁺T cells (1 week HFHS diet) with or without DTX-mediated Treg depletion. n = 4–6 biological replicates per group from two independent experiments. Depicted are Box-and-whisker plots (min-to-max values with single data points). Two-tailed unpaired t test, P(b) = 0.0040; P(c) = 0.0014; P(d) = 0.0440; P(e) = 0.3135; P(h) = 0.0124; P(i) = 0.065; P(j) = 0.0272; P(l, hypothalamus) = 0.0008; P(l, rest brain) = 0.0001; P(m, hypothalamus) = 0.0346; P(m, rest brain) <0.0001; Source data are provided as a Source Data file. *P < 0.05; **P < 0.01; ****P < 0.0001.