Postnatal 0 to 3 day WT or Trem2−/− mice were used to prepare primary microglia. The procedure has been previously described (43). After 14 days of culture, primary microglia were collected and used for experiments. For small interfering RNA (siRNA) transfection, primary microglia were seeded overnight and then transfected with 50 nM siRNA using Lipofectamine RNAiMAX (Invitrogen, Waltham, MA, USA). Aβ phagocytosis assays were performed at 48 to 72 hours after transfection. siRNA sequences used are listed as follows: (i) Trem2 #1: uucuccugagcaaguuucuug, Trem2 #2: caucacucugaagaaccucca; (ii) CR3 #1: aucuuuccugcuaauucugag, CR3 #2: ccugcuaauucugaggucaca; (iii) GPR34 #1: auaaccaccaagcaaaguauu, GPR34 2#: cucauggaaugcacuuuauaa; (iv) MerTK #1: guuuaaucacaccauuggaca, MerTK #2: agauguugugauugacagaaa.

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