Parasite Propagation and Atovaquone Treatment

For the experiments described here, we used MRC5 parasite line previously characterized [21 (link)] and its parental strain, RHΔhpt+GFP+βgal, which expresses GFP and β-galactosidase (βgal) [62 (link)]. Toxoplasma tachyzoites were propagated by passaging in human foreskin fibroblasts (HFFs, purchased from the American Tissue Culture Collection, ATCC) in a humidified incubator maintained at a temperature of 37°C and 5% CO₂ concentration. Normal growth medium used was Dulbecco’s Modified Eagle Medium (Life Technologies) supplemented with 10% fetal bovine serum (Atlanta Biologicals), 2 mM L-gluatamine (Life Technologies) and 50 μg/mL penicillin-streptomycin (Life Technologies). Atovaquone was purchased from Sigma-Aldrich and prepared using DMSO as solvent.