Neuronal response to ApoE isoforms

Cerebral cortices were harvested from newborn (P0–P2) C57BL/6 mice as previously described^{15 (link)}. Neurons were cultured with 150 nM human recombinant ApoE3 (PeproTech 350-02, NJ, USA) or ApoE4 (PeproTech 350-04) combined with 100 μM PA for 24 h. In some experiments, neurons were preincubated with 10 μM capsaicin for 30 min followed by the addition of ApoE3 or ApoE4 and PA. Human neuroblastoma-derived SH-sy5y cells were exposed to 150 nM human recombinant ApoE3 or ApoE4 combined with 100 μM PA for 24 h. Microglia-conditioned medium (MCM) was collected from the culture medium of human recombinant ApoE3- and ApoE4-treated BV2 cells. BV2 cells were preincubated with 10 μM capsaicin for 30 min, and then ApoE3 or ApoE4 was added. DIV8 neurons were incubated with 95% neuronal medium and 5% MCM at 37 °C with 5% CO₂ for 24 h.