FIG 3.

Quantification of BAG1 and SAG1 gene expression during treatment with BKI-1294. Confluent HFF monolayers grown in 6-well plates were infected with 2.5 × 10⁴ tachyzoites of T. gondii RH overexpressing CDPK1_G128M or wild-type CDPK1. Treatments with 5 μM BKI-1294 or DMSO (control) were initiated 2 days postinfection. After different time points, the adherent cells were harvested for RNA extraction. mRNA levels of BAG1, SAG1, and GRA2 were quantified by real-time RT-PCR. Mean values (±SE) from four replicates were assessed, and expression levels of BAG1 (bradyzoite specific) or SAG1 (tachyzoite specific) relative to the amount of GRA2 (overall) mRNA were given as multiples of the corresponding DMSO control values (MV ± SE for quadruplicates).