Fig. 2: Validating screen results with diCRE mediated excision.

a, b TK-diCRE (shown in green) and ribonucleotide reductase (RNR) -diCRE (shown in purple) parasites were generated and used to infect an HCT8 monolayer in the presence or absence of rapamycin. Genomic DNA was extracted at 6, 24, and 48 h, and diagnostic PCRs confirmed the level of excision at the given time points. Data shown is representative of 2 biological replicates. c HCT8 cell monolayers infected with TK-diCRE and RNR-diCRE parasites in the presence or absence of rapamycin. At 6, 24, and 48 h the monolayers were fixed, stained, and the number of parasites per host nuclei was quantified. Box plot whiskers show the minimum and maximum values, the box shows the 25th to 75th percentile and the line shows the median. Representative images are shown, with nuclei in blue (Hoechst), and parasites in green (Vicia villosa lectin). Scale bar = 30 μm. Data shown is representative of 2 biological replicates. Significance was determined using a two-tailed unpaired t test. d TK-diCRE or RNR-diCRE parasites were used to infect Ifnγ^–/– mice, which were either treated with rapamycin or vehicle (DMSO) in their drinking water starting at day 2 post infection. Data shows the mean faecal luminescence from a pooled cage sample ( ± SEM of 2 technical replicates), n = 2 mice per condition. Note that a cell culture dish in the figure indicates an in vitro experiment, while a mouse silhouette indicates an in vivo experiment.