Extended Data Fig. 4: Defects of the ∆TgVIP1 strain.
(a) Plaque assays of parental and ∆TgVIP1 strains showing representative images and quantification of the lysed area from 3 independent experiments. Insets show summary plots of the mean number of plaques or plaque area for the 3 independent experiments. Data are means ± SD. p values for number of plaques: p = 0.0384 (exp 1); p < 0.0001 (exp 2); p = 0.0164 (exp 3); Mean number of plaques p = 0.0052; for plaque area: p = 0.0023 (exp 1); p = 0.0008 (exp 2); p = 0.0048 (exp 3); Mean plaque area: p = 0.0059 (unpaired two-tailed t test) (b) Uracil incorporation assays of uninfected and HFF-1 cells infected with parental and ∆TgVIP1 strains for 24-h. Shown are 3 independent assays and in the inset the mean percent of parental uracil incorporation from all 3 experiments. Data are means ± SD. *** p < 0.0001; ** p = 0.0003 (One-way ANOVA with a Tukey’s multiple comparisons test (exp 1, exp 2, and exp 3); unpaired two-tailed t test (for percent of parental) (c) EM of HeLa or HFF-1 cells infected with ∆TgVIP1 parasites for 24-h. The boxed region is shown in the enlargement. Black arrowheads denote the aberrant e-dense structures containing vesicles and the white arrowheads denote the less e-dense aberrant structures containing fibrils. (d) EM of HeLa cells infected with ∆TgVIP1 parasites for 6-h, showing the host ER coverage of the PV. The boxed region is shown in the enlargement. hcell, host cell; P, parasite; hm, host mitochondria; Go, parasite Golgi; m, parasite mitochondrion. (e) HeLa WT cells were infected with WT parasites or Δvip1 for 6-h, 24-h, or 48-h, fixed, and processed for EM. The percent coverage of the PV by the host ER was measured. Data are means ± SD. * p = 0.0007 (24-h); p = 0.0438 (48-h) (unpaired two-tailed t test); number of PV (all samples and times) = 19.
