Figure S3.
G795A microglia proliferate along distinct wavefronts in the murine brain. (A) Representative confocal 10× stitch image of engrafted and proliferating G795A microglia after 30 d of CSF1Ri. Ki67+ mitotic cells (blue), Ku80+ human nuclei (red), and IBA1+ microglia (green). (B) 40× representative images of Ki67+ G795A microglia in proliferating wavefront and Ki67− G795A microglia within the initial hippocampal injection site. (C) Proliferating microglia in the wavefront exhibit significantly higher levels of human nuclear marker Ku80. Unpaired t test t(4) = 3.008, *P = 0.0396. (D) Representative 10× stitch image of P2RY12 expressing G795A microglia after 30 d of CSF1Ri. P2RY12 (purple), Ku80 (red), IBA1 (green). (E) 40× representative images of P2RY12-low G795A microglia in the wavefront and P2RY12-high G795A microglia within the hippocampus. (F) Proliferating microglia in the wavefront exhibit significantly lower levels of microglia homeostatic marker P2RY12. Unpaired t test t(4) = 7.545, **P = 0.0017. (G) Representative 10× stitch image of LGALS-3 expressing G795A microglia after 30 d of CSF1Ri. LGALS-3 (orange), Ku80 (red), IBA1 (green). (H) 40× representative images of G795A microglia expressing LGALS-3 within the proliferative wavefront with diminished expression within the hippocampus. (I) Proliferating microglia in the wavefront exhibit significantly higher levels of microglia activation marker LGALS-3. Unpaired t test t(4) = 9.511, ***P = 0.0007. Data represented as integrated density normalized to number of Ku80+/IBA1+ human microglia per FOV for all antibodies calculated from three matched coronal sections per animal (n = 3 biological replicates per region). Error bars, SEM. Representative 10× stitch scale bar, 100 µm. Representative 40× scale bar, 25 µm.

G795A microglia proliferate along distinct wavefronts in the murine brain. (A) Representative confocal 10× stitch image of engrafted and proliferating G795A microglia after 30 d of CSF1Ri. Ki67+ mitotic cells (blue), Ku80+ human nuclei (red), and IBA1+ microglia (green). (B) 40× representative images of Ki67+ G795A microglia in proliferating wavefront and Ki67− G795A microglia within the initial hippocampal injection site. (C) Proliferating microglia in the wavefront exhibit significantly higher levels of human nuclear marker Ku80. Unpaired t test t(4) = 3.008, *P = 0.0396. (D) Representative 10× stitch image of P2RY12 expressing G795A microglia after 30 d of CSF1Ri. P2RY12 (purple), Ku80 (red), IBA1 (green). (E) 40× representative images of P2RY12-low G795A microglia in the wavefront and P2RY12-high G795A microglia within the hippocampus. (F) Proliferating microglia in the wavefront exhibit significantly lower levels of microglia homeostatic marker P2RY12. Unpaired t test t(4) = 7.545, **P = 0.0017. (G) Representative 10× stitch image of LGALS-3 expressing G795A microglia after 30 d of CSF1Ri. LGALS-3 (orange), Ku80 (red), IBA1 (green). (H) 40× representative images of G795A microglia expressing LGALS-3 within the proliferative wavefront with diminished expression within the hippocampus. (I) Proliferating microglia in the wavefront exhibit significantly higher levels of microglia activation marker LGALS-3. Unpaired t test t(4) = 9.511, ***P = 0.0007. Data represented as integrated density normalized to number of Ku80+/IBA1+ human microglia per FOV for all antibodies calculated from three matched coronal sections per animal (n = 3 biological replicates per region). Error bars, SEM. Representative 10× stitch scale bar, 100 µm. Representative 40× scale bar, 25 µm.

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