Primary neurons infections and bradyzoite in vitro targeting assay

Neo-natal rat brain cells were collected from hippocampus and seeded as described in reference (22). After 14 days of maturation, the primary brain cells were infected using 5,000 parasites, of the PTG-LUC-GFP strain, per well of a 24-well plate at a MOI of 0.1. Fourteen days later, the infected primary cell culture was incubated with a compound (MC2590, MC2059, or MC2125) concentration of three times the IC₅₀ (as determined above against the tachyzoite stage) or an equivalent volume of DMSO. The culture was left for 7 more days, then collected and lysed in 100 µL of lysis buffer, and processed as described before for luciferase detection.