Ham s f 12 nutrient mix

Primary macrophage cultures were generated from bone marrow via differentiation in 40 ng/ml M-CSF, as described (Samuel et al., 2006 (link)). Embryonic fibroblasts were generated and maintained as previously described (Szretter et al., 2012 (link)). Primary cultures of cerebral cortical neurons and purified cerebellar granule cell neurons were generated using E15 embryos or P3 neonatal mice, as described (Klein et al., 2005 (link)). Purified microglia were obtained from mixed glial cultures generated from P1-P3 neonatal mice as described (Williams et al., 2014b (link)). Human neuroblastoma cell lines NB8, NB15, NB16 were maintained in a 1:1 mixture of DMEM and Ham’s F-12 Nutrient Mix (ThermoFisher). Neuroblastoma cells were differentiated into neuron-like cells by supplementing culture medium with 10μM all-trans retinoic acid (Sigma-Aldrich) and 50ng/ml recombinant human BDNF (Peprotech), as described (Encinas et al., 2000 (link)). For infection experiments, macrophage and cultures were infected at MOI 0.01, while primary neuronal cultures and neuroblastoma cell lines were infected at MOI 0.001.

The female human embryonic kidney cell line HEK293T (Cat#CRL-3216; RRID: CVCL_0063) was obtained from the American Type Culture Collection (ATCC) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) with 4.5 g/L glucose, L-glutamine, and sodium pyruvate (Corning) supplemented with 10% (v/v) fetal bovine serum (Avantor Seradigm) and Penicillin-Streptomycin (Gibco). Immortalized mouse hippocampal cell line mHippoE-2 (Cat# CVCL_D377; RRID: CVCL_D377) was obtained from Cedarlane Laboratories and cultured in DMEM with 4.5 g/L glucose, L-glutamine, and sodium pyruvate (Corning) supplemented with 10% (v/v) fetal bovine serum (Avantor Seradigm) and Penicillin-Streptomycin (Gibco). The sex of mHippoE-2 cells was not disclosed by the manufacturer and may be a mixed population of male and female. The female human neuroblastoma cell line SH-SY5Y (Cat#CRL-2266; RRID: CVCL_0019) was obtained from the ATCC and cultured in equal parts Minimum Essential Medium Eagle (EMEM) with Earle′s salts, L-glutamine and sodium bicarbonate (Sigma-Aldrich) and Ham’s F12 Nutrient Mix (ThermoFisher), supplemented with 10% (v/v) gamma-irradiated and heat-treated fetal bovine serum (Avantor Seradigm) and Penicillin-Streptomycin (Gibco). Cell lines used in this study have not been authenticated. Cell lines used in this study were cultured in a humidified 37°C incubator with 5% CO₂.