Extended Data Fig. 4: Primary microglia engulf synaptosomes through extrinsic SPP1. (a) Schematic ill...

Primary microglia engulf synaptosomes through extrinsic SPP1. (a) Schematic illustrating in vitro engulfment assay whereby primary microglia isolated from WT or Spp1 KO/KO mice phagocytose S26C oAβ-treated synaptosomes that are tagged with pHrodo upon PBS or recombinant SPP1 treatment. (b) Representative images showing engulfment of pHrodo-synaptosomes by primary microglia isolated from WT mice (upper panel) or Spp1 KO/KO mice (lower panel), treated with vehicle (left) or 100 nM recombinant SPP1 (right). Data are representative of primary microglial cultures prepared from n = 6−8 neonates per genotype, examined over 2 independent experiments. Scale bar represents 100 µm. (c) Representative FACS plot showing pHrodo signal in CD11b + cells collected from primary microglial culture (upper). No pHrodo signal was observed after Bafilomycin treatment, confirming lysosomal acidification as the source of pHrodo signal (lower). Data are representative of 2 independent experiments.

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