S1 Fig -

A. Schematic representation of the strategy used to generate TgCDPK7-iKD parasites. A plasmid construct that allowed insertion of transactivator TATi-1 and replacement of TgCDPK7 promoter with the 7tet-Op SAG1 (TetO7) inducible promoter was introduced by double homologous recombination in TgCDPK7 locus. After transfection, the drug selected parasites were cloned by limiting dilution. B. Genotyping of TgCDPK7-iKD parasites. Although two independent clones were obtained, clone#1 (TgCDPK7-iKD) was used for most reported studies and some experiments were performed with clone #2 (Panel E). PCR amplification of the unmodified and recombined locus was performed using indicated primers (Panel A, S1 Table ) for confirming 5’- and 3’- integration. PCR products of expected size were obtained and the wild type (WT) locus was absent in the transgenic TgCDPK7-iKD parasites. C. TgCDPK7-iKD parasites grown in the absence or presence of ATc for 72h. Real time PCR was performed for assessing the expression of TgCDPK7. TgCDPK3 was used as a control with respect to which TgCDPK7 expression was determined (Mean+/-SE, *n = 3, p<0.0001, t-test). D. ΔKu80 parasites were pre-incubated for 48h with ATc and were subsequently allowed to invade fresh HFFs in the presence or absence of ATc. The number of parasites per vacuole was determined after 24h. ATc treatment did not alter replication of ΔKu80 parasites. There was no significant difference in parasite growth upon ATc treatment. E. TgCDPK7-iKD_clone 2 was used for performing parasite replication assays in the presence or absence of ATc and ethanolamine as described in Fig 5D . (mean±SE, *n = 3, p<0.01 for 8 parasites/vacuole, ANOVA). (PDF)