miRNA-Seq of EVs from mus musculus primary cortical neurons infected with T. gondii: EVs from Infected neurons, replicate 2
Primary cortical murine neurons were plated 5.0x10^5 cells per well in 2 mL in 6 well plates. Eight days after culture neurons were infected with T. gondii at an MOI (multiplicity of infection) of 0.50. Neuronal supernatant was collected and filtered through a 0.8 um filter, 9 days and 11 days after culture. Supernatant were spun at 500 g for 15 minutes at 4C and then again at 15,000 g for 20 minutes at 4C. EV miRNA was isolated with the exoRNAeasy Midi Kit (Qiagen, 77144) according to the manufacturer's protocol. RNA was harvested using Rneasy mini kit (Qiagen).