Figure 4.

MHC class II^low macrophages are a female-specific subset with restricted localization

(A) scRNA-seq analysis of CD74, H2-Aa, Ciita, and the KEGG pathway “Antigen processing and presentation” expression among myeloid cells.

(B) (Top) Flow-cytometry plots showing F4/80 and MHC class II expression among AGMs from 7-week-old male and female mice. (Bottom) Proportions and numbers of MHC class II^high and class II^low AGMs from 7-week-old male and female wild-type mice. ♂ n = 16 and ♀ n = 15. Data are pooled from 3 independent experiments.

(C) Fluorescence-microscopy analysis of CD68 and MHC class II expression in adrenal glands from 7-week-old male and female mice. Scale bar: 200 μm. M, medulla; C, cortex. Data are representative of at least 3 independent experiments.

(D) Distribution of CD68⁺ cells between cortex and medulla from adrenal glands of 7-week-old male and female mice. Data are represented as proportion of CD68⁺ cells from each zone among total cells. ♂ n = 4 and ♀ n = 6. Quantification from one experiment.

(E) Proportion of MHC-II^high and MHC-II^low CD68⁺ cells in the cortex and medulla of adrenal glands from 7-week-old male and female mice. ♂ n = 4 and ♀ n = 6. Quantification from one experiment.

(F) Analysis of macrophage metabolic activity using SCENITH, represented by Puromycin MFI (n = 14). Data pooled from 4 independent experiments.

(G) Measure of glycolytic and mitochondrial metabolism in macrophages using SCENITH (n = 10–11). Data pooled from 3 independent experiments.

Statistical analysis was performed using two-tailed Mann-Whitney tests (panel B, quantifications), two-way ANOVA with Bonferroni’s post-test (proportions in panel B, panel D and panel E), or paired Wilcoxon t-tests (panels F and G). See also Figure S4.