Fig. 3: Cytokine-controlled neutrophilia depends on the AR in BM stroma.
a, b Mice were injected i.p. twice daily with 125 µg/kg of G-CSF or vehicle on three consecutive days. Neutrophil (Ly6G+) counts in blood were measured by flow cytometry; representative flow plots in a. a Vehicle or G-CSF injection in control or O-ARKO mice; *P = 0.034 (two-sided unpaired Student t test); n = 3 + 5 and 14 + 14 for vehicle and G-CSF respectively. b G-CSF injection in sham-castrated (control) or castrated (ORX) mice; *P = 0.016 (two-sided Mann–Whitney); n = 4 + 5. c, d Cxcl12 expression was analyzed by qPCR in CD45- (CD45/TER119 depleted) bone cells from O-ARKO or control mice (c) and CD45− (CD45+ depleted) bone marrow (BM) cells from ORX or control mice (d). c *P = 0.031 (two-sided unpaired Student t test); n = 16 + 16. d *P = 0.037 (two-sided unpaired Student t test); n = 13 + 13. e BM cells from Osterix-Cre+ tdTomato reporter mice were cultured for 10 days, tdTomato+ and tdTomato- cells were sorted and Cxcl12 expression analyzed by qPCR; **P = 0.0018 (two-sided unpaired Welch’s t test); n = 4 + 4. f–k Analysis of Ar and Cxcl12 distribution in a single cell RNA sequencing dataset of mouse stromal cells (ref. 32). f In the overview of 18 stromal cell clusters colored by clustering (UMAP visualization), clusters 2 and 10 are multipotent mesenchymal stem/stromal cells (MSC) and cluster 11 osteolineage (OL) cells. g–j The expression intensity and distribution of Ar, Cxcl12, Lepr, and Bglap RNA in stromal cell clusters. k Percentages and expression intensity of Ar and Cxcl12 in stromal cell clusters. l, m BM stromal cells from C57BL/6 J male mice were stimulated with the AR agonist dihydrotestosterone (DHT) or vehicle for 10 days. l Cxcl12 mRNA expression; *P = 0.028 (two-sided unpaired Student t test); n = 6 + 6. m CXCL12 protein levels in the cell supernatant; *P = 0.048 (two-sided Mann–Whitney); n = 6 + 6. Bars indicate means (a, c–e, l) or medians (b, m), error bars are SEM (a, c–e, l) or interquartile ranges (b, m), and circles represent individual mice. Source data are provided as Source data file.
