ΔdblGATA mice have reduced monocyte numbers during EAE. WT and ΔdblGATA mice were immunized for EAE induction. Ten mice per group were initially immunized, and 3 to 4 mice were pooled for each data point when the mice were euthanized on day 16 after immunization. Representative data from 4 experiments are shown. (A) Numbers of CD45⁺ cells isolated from the CNS (spinal cords and brains combined) of WT and ΔdblGATA mice with EAE. (B) Flow cytometry plots depicting CD45 expression vs side scatter, and quantification of CD45^hi and CD45^lo cells among the total cells isolated from the CNS. (C) Frequency of CD11b⁺CD11c⁺ and CD11b⁺Ly6C^hi cells among the CD45⁺ cells. (D) t-distributed stochastic neighbor embedding (t-SNE) plots of CD45⁺ cells. Monocyte-derived DCs (MDCs) were defined as CD45^hiCD11b⁺Ly6G^lo/−CD11c⁺Ly6C⁺MHC-II⁺. Undifferentiated monocytes (Mo) were defined as CD45^hiCD11b⁺Ly6G^lo/−Ly6C^hiMHC-II⁻. Microglia were defined as CD45^loCD11b⁺. Neutrophils were defined as CD45⁺CD11b⁺Ly6G⁺Ly6C^intermediate. CD4⁺ cells were defined as CD45^hiCD11b⁻CD4⁺. Other antigen-presenting cells (APCs) were defined as CD45^hiCD11b⁺Ly6G⁻MHC-II⁻Ly6C⁻. The other CD45⁺ cells were CD45⁺CD11b⁻CD4⁻CD11c⁻. (E) Heatmap overlay of CD11c, MHC-II, CD80, and CD86 expression. (F) Flow cytometry plots showing CXCL9 and arginase 1 (Arg1) expression among CD45⁺Sall1⁻CD11b⁺CCR2⁺Ly6C^hi monocytes. (G) Quantification of the frequency of CXCL9⁺, arginase1⁺, pro–IL-1β⁺, TNF⁺, and IL-10⁺ cells among CD45⁺Sall1⁻CD11b⁺CCR2⁺Ly6C^hi monocytes. For panels F and G, n = 9 to 10 mice per group. Significance was determined by 2-tailed unpaired Student t test. Error bars are standard error of the mean (SEM). MHC-II, major histocompatibility complex II; TNF, tumor necrosis factor.