Supplementary Figure 1: Sall1 expression is restricted to microglia within the hematopoietic system.

(a) Flow cytometry plots show representative pre-gating strategy for CD45⁺ cells (shown are CNS cells). (b) Flow cytometry analysis of GFP (Sall1) expression in different organs of Sall1^GFP/+ and Sall1^+/+ (control) mice (pre-gated on CD45⁺ cells as in a. (c) qPCR analysis of Sall1 mRNA in sorted cell populations derived from WT mice; results were normalized to Pol2 expression. Alveolar MFs: CD45⁺Siglec-F⁺CD11c⁺, Lung CD11b⁺ DCs: CD45⁺Siglec-F⁻CD11c⁺MHCII⁺CD11b⁺, Lung CD103⁺ DCs: CD45⁺Siglec-F⁻CD11c⁺MHCII⁺CD103⁺, SP MF (spleen macrophages): F4/80^hiCD11b⁺, SP NPs (spleen neutrophils): Ly6G⁺SSC^hi, BM Mo (BM monocytes): Lin⁻CD11b⁺Ly6C⁺CD115⁺, microglia: CD45^loLy6C⁻Ly6G⁻CD11b⁺F4/80⁺, Per. B cells (peritoneal B cells): B220⁺, Per. (peritoneal) MFs: CD115⁺CD11b⁺F4/80⁺. (d) qPCR analysis of Sall1 mRNA in total tissue lysates of different organs; results were normalized to Pol2 expression. (e) Representative flow cytometry plots of kidney, liver and heart of Sall1^GFP/+ mice (pre-gated on CD45⁻ cells). (f) Quantification of results in e, presented as frequency of GFP⁺ (Sall1) cells; each symbol represents an individual mouse; small horizontal lines indicate the mean (± s.e.m.). Data are representative of 2-4 mice per genotype, 2 experiments (b); 2 samples per population pooled from 2-3 WT mice, 2 experiments (c; mean ± s.e.m.); 13 (spleen), 12 (brain), 11 (kidney, liver), 9 (spinal cord), 7 (lung, heart), 4 (skin), 3 (lymph node) WT mice, 2-5 experiments, 1 experiment (lymph node) (d; mean ± s.e.m.); 6 (liver), 5 (kidney, heart) Sall1^GFP/+ mice, 2 experiments (e,f).