Supplementary Figure 3: CNS-infiltrating myeloid cells and BM-derived microglia and/or macrophages do not express Sall1.

(a) Frequency of GFP⁺ microglia and CD45^hi MF of Sall1^GFP/+ and Cx3cr1^GFP/+ reporter mice and of YFP⁺ or RFP⁺ microglia and CD45^hi MFs in tamoxifen treated Sall1^CreERR26-YFP or Cx3cr1^CreERR26-RFP mice. Microglia: CD45^loCD11b⁺F4/80⁺Ly6C⁻Ly6G⁻, CD45^hi MFs: CD45^hiCD11b⁺F4/80⁺Ly6C⁻Ly6G⁻. (b) Flow cytometry analysis and quantification of the frequency of GFP⁺ microglia in tamoxifen- and diphtheria toxin-treated Sall1^+/+(CD45.1)→Cx3cr1^CreER-iDTR(CD45.2) or Sall1^GFP/+(CD45.1)→Cx3cr1^CreER-iDTR(CD45.2) BM chimeras on day 0, 11 and 21 after treatment (pre-gated on CD11b⁺F4/80⁺CD45.1^loLy6C⁻Ly6G⁻ cells) or in untreated Sall1^GFP/+ (control) mice. (c) Representative histograms and quantification of GFP expression in monocyte-derived cells (MCs) (gated on CD45^hiCD11b⁺CD11c⁺MHCII⁺Ly6C⁺), neutrophils (gated on CD45^hiCD11b⁺ Ly6G⁺), CD4⁺ T cells (gated on CD45^hiCD11b^-CD4⁺) and microglia (gated on CD45^loLy6C^-Ly6G^-CD11b⁺) in Sall1^GFP/+ mice at peak disease of MOG_35-55/CFA-induced EAE. Each symbol (a-c) represents an individual mouse; small horizontal lines (a-c) indicate the mean (± s.e.m.). *P < 0.0001 (one-way ANOVA). Data are representative of 15 (Sall1^GFP/+), 10 (Sall1^CreERR26-YFP, Cx3cr1^CreERR26-RFP), 4 (Cx3cr1^GFP/+) mice, at least 2 experiments (a); 4 mice (d11, d21), 2 experiments and 1 mouse (d0, untreated Sall1^GFP/+) (b); 7 (microglia), 5 (MCs, Neutrophils, CD4⁺ T cells) mice, 3 experiments (c).