Figure 2.

scRNA-seq analysis reveals adrenal gland leukocyte diversity and monocyte contribution to the macrophage pool

(A) scRNA-seq analysis of adrenal gland CD45⁺ cells from 7-week-old male and female wild-type mice.

(B) Proportion of each cluster identified in scRNA-seq analysis.

(C) Heatmap showing normalized expression levels of cluster-specific genes.

(D) Violin plots showing Cx3cr1, Timd4, and Lyve1 expression by cells from clusters 1–6.

(E) Flow-cytometry plot showing Ly6C and CCR2^GFP expression among adrenal gland CD45⁺CD11b⁺CD64⁺ cells in male and female CCR2^GFP/+ mice. Data are representative of three independent experiments.

(F) Flow-cytometry plot showing Ly6C and TdTomato expression among adrenal gland CD45⁺CD11b⁺CD64⁺ cells from male and female CCR2^creERT2 x R26^TdTomato mice 48 h after TAM gavage. Data are representative of two independent experiments.

(G) Quantification of TdTomato⁺ macrophages in 16- to 20-week-old male and female heterozygous (CCR2^+/-, ♂ n = 6 and ♀ n = 6) or double knockin (CCR2^-/-, ♂ n = 4 and ♀ n = 3) CCR2^creERT2 x R26^TdTomato mice 48 h after TAM gavage. Data are pooled from two independent experiments.

(H) Histograms representing R26^TdTomato expression in AGMs from 10-week-old female CCR2^creERT2/+ x R26^TdTomato mice 2, 7, and 14 days after TAM gavage. Data are representative of one (days 7 and 14) or two (day 2) experiments.

(I) Proportions of TdTomato⁺ macrophages from 10-week-old female CCR2^creERT2/+ x R26^TdTomato mice 2 (n = 8), 7 (n = 4), and 14 (n = 3) days after TAM gavage. Data from one (days 7 and 14) or two (day 2) experiments.

(J) Proportions of TdTomato⁺ macrophages from 8 (♂ n = 2, ♀ n = 6) or 16 (♂ n = 5, ♀ n = 5)-week-old male and female Ms4a3^cre/+ x R26^TdTomato mice. Data are from one experiment.

(K) Quantification of AGMs in male and female CCR2^+/- (♂ n = 18, ♀ n = 21) and CCR2^-/- (♂ n = 10, ♀ n = 10) mice. Data are pooled from four independent experiments.

(L) Proportions of Ki67⁺ AGMs in male and female CCR2^+/- (♂ n = 11, ♀ n = 11) and CCR2^-/- (♂ n = 6, ♀ n = 5) mice. Data are pooled from two independent experiments.

Statistical analysis was performed using two-way ANOVA with Bonferroni’s post-test.

See also Figure S2.