Figure 2.

Quantification of replication inhibition using replication assay. Compounds were tested for replication inhibition using established procedures.²⁴ HFF monolayers were inoculated with tachyzoites and then incubated (37 °C, 5% CO₂) for 2 h. Compounds (final 10 μM) or DMSO (vehicle) were then added to the medium. Parasite replication proceeded for 24-26 h after which time the monolayers were fixed, permeablilized, and immunolabeled with Rb anti-p30 followed by Gt antirabbit Alexa Fluoro 594 (red). DAPI was added to secondary antibody. Cells were examined by reflected fluorescence as in Figure 1. Data were compiled from three independent experiments, each from 10 random fields per sample. The number of vacuoles containing 1, 2, 4, or 8+ parasites/vacuole were enumerated by eye. Data are expressed as the modal number of parasites per vacuole.