figs2RT-qPCR verification of RNA-Seq results. Expression of trg, TGGT1_311100, and bag1 transcripts in RNA extracted from T. gondii parasites treated with trtE (24.2 nM, red) or DMSO carrier control (0.1%, black) at 1, 2, and 4 h post treatment. Fold-change in gene expression was calculated using the 2^−ΔΔCt method with three replicates. Treatment of parasites with trtE significantly increased the expression of trg at all time points (p = 0.04, 0.01, and 0.004 at 1, 2, 4 h, respectively). Expression of TGGT1_311100 during trtE treatment was not significantly different from treatment with DMSO (p = 0.4, 0.5, and 0.7 at 1, 2, 4 h respectively). At 4 h post treatment with trtE, expression of bradyzoite-specific gene bag1 was significantly lower (p = 0.001) than DMSO-treated parasites. Three technical replicates were performed on all samples.