Fig. 1.

Gating strategies and detection of LAG-3 expression by FITC conjugated LAG-3 antibody. A Gating strategy of isolating single CD11⁺/CD45^low cells from the brains of PBS injected mice. Single-cell suspension from the brains of naïve mice involved pre-gating by forward scatter area (FSC-A) vs. side scatter area (SSC-A) plot, and forward scatter height (FSC-H) vs. FSC-A density plot to select singlets. Microglia were then identified by the high expression of CD11b and the low expression of CD45. B Distribution of fluorescence intensities from FITC conjugated LAG-3 antibody and isotype control antibody treated microglia. C Gating strategy of isolating single CD11⁺/CD45^low microglia cells and CD11⁻/CD45^high lymphocytes from the brains of LPS injected mice. D, E Distribution of fluorescence intensities from FITC conjugated LAG-3 and isotype control antibody-treated microglia (D) and lymphocytes (E) from LPS injected mice. All data were replicated in four independent experiments