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. Author manuscript; available in PMC: 2020 Jan 1.
Published in final edited form as: Mol Microbiol. 2018 Nov 28;111(1):46–64. doi: 10.1111/mmi.14138

Figure 4. Establishment and growth of parasite strain with conditional expression of TgDrpC.

Figure 4.

A) Western blots of extracts from parasites in which the endogenous TgDrpC was tagged with either HA or HA-DD grown for 24 or 48 hours in presence (+) or absence (-) of Shld1. Blots were probed with antibodies against HA to detect TgDrpC or against the surface antigen SAG1 as a loading control. B) Parasites expressing TgDrpC-HA-DD were allowed to grow for 5 and 8 days in the presence and absence of Shld1. Cultures were stained with Crystal Violet, which allows visualization of plaques (white areas) arising from parasites lysing their host cells (dark areas). C) Doubling assays quantifying the number of TgDrpC-HA-DD parasites per vacuole following 24 and 48 hours of growth in the presence or absence of Shld1 (n=3, ±SD). D) The total number of vacuoles of TgDrpC-HA-DD parasites at 24 and 48 hours with and without Shld1 was determined for 10 randomly selected fields of view. (n=3, ±SD) (***p<0.001). E) Phase image of TgDrpC-HA-DD parasites grown without Shld1 for 48 hours reveals aberrant vacuole structures. F) The distribution of vacuoles that appear normal, swollen, or vacuolated when parasites are grown with or without Shld1 for 48 hours (n=3, ±SD).