G-CSF induces emergency granulopoiesis during OPC.

(A) Csf3 mRNA expression in the tongues of naïve (n) and infected (inf) WT mice was quantified by qRT-PCR 24 hours post-infection. (B) Total numbers of CD45⁺ cells in the bone marrow of naïve (n) and infected (inf) WT mice were assessed by flow cytometry. (C) Representative FACS plots showing the analysis of Ly6G^hi CD11b⁺ Ly6C^int mature neutrophils and Ly6G^lo CD11b⁺ Ly6C^int immature neutrophils in the bone marrow of naïve and infected WT mice. Depicted events were pre-gated on singlets, scatter, CD45⁺ alive cells. Numbers indicate the % of cells in each gate. (D—E) Analysis of Ly6G^hi mature neutrophils (D) and Ly6G^lo immature neutrophils (E) in the bone marrow of naïve (n) and infected (inf) WT mice 24 hours post-infection. (F–G) As in D–E, but mice were treated with anti-G-CSF or left untreated prior to infection as indicated. (H–J) WT mice were treated with anti-Ly6G and/or anti-G-CSF antibody or left untreated prior to infection as indicated. CD45⁺ CD11b^hi Ly6C^int neutrophils were quantified in the blood (H) and tongues (I) 24 hours post-infection. The fungal burden in the tongue was determined on day 3 post-infection (J). Each symbol represents an individual mouse and the lines represent the mean (B, D–G) or geometric mean (A, H—J) of each group. Data are pooled from two (F, G) or representative of two independent experiments (A–E, H–J). Statistical analysis was performed using log₁₀ transformation (A, H–J) and Student’s t-test with Welch’s correction (A–B, D–E) or a one-way ANOVA with Dunnett’s test (F–J).