Fig. 3

Assessment of phagocytosis in circulating monocytes and MDMs in the brain. a Experimental timeline. Asplenic mice were received infusion of GFP⁺ splenocytes and bead^580/605 at 6 days after MCAO and 2 days after splenectomy (SPX). Following the infusion, blood and brain tissue were collected at several time points up to 24 h, as indicated by arrows. b Assessment of bead⁺/GFP⁺ population in circulating monocytes at baseline at 0 h (pre-infusion of GFP splenocytes and bead^580/605; Pre) as well as at 5 m, 15 m, 30 m, 4 h, and 24 h post-infusion. Lin (lineage markers) is the mixture of antibodies against T cells (CD90.2), B cells (CD45R/B220), natural killer cells (NK1.1), and granulocytes (Ly6G). The monocyte population was identified by CD11b⁺/Lin⁻ cells (indicated by black boxes) and were further gated for GFP⁺/bead⁺] population (indicated by red boxes). c Assessment of MDMs [GFP⁺/bead⁺/CD11b⁺/CD45⁺] in the post-ischemic brain 30 m, 4 h, and 24 h following infusion of GFP splenocytes and bead^580/605. MDMs are in the boxed region. Representative plots from three independent experiments per time point. d Immunofluorescence images of the striatum taken at 7 days post-stroke with arrows (white) indicating phagocytic MDMs [GFP⁺/bead⁺/Iba1⁺], arrowheads indicating either microglia or infiltrated endogenous non-tagged MDMs [GFP⁻/bead⁺/Iba1⁺], and * (aqua) indicating non-phagocytic MDMs [GFP⁺/bead⁺/Iba1⁺]. Scale bar = 100 µm