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. 2017 Feb 28;8(1):e02189-16. doi: 10.1128/mBio.02189-16

FIG 5 .

FIG 5 

TgDJ-1 regulates microneme secretion. (A) Western blots of microneme secretion assays comparing levels of basal secretion of MIC2 over time for wild-type (WT), ΔTgdj-1 clone F6, and ΔTgdj-1 parasites complemented with wild-type Tgdj-1 (ΔTgdj-1WTcomp). GRA7, loading control. (B) Quantification of basal microneme secretion assays; data represent means ± SD for results from n = 3 experiments. Significance was calculated for a two-way statistical ANOVA for comparison of the different strains (WT, ΔTgdj-1, and ΔTgdj-1WTcomp) (****, P ≤ 0.0001) and the different time points (15, 30, and 60 min) (****, P ≤ 0.0001). Post hoc pairwise comparisons of the different strains at each time point were performed using Tukey’s multiple-comparison test and the data presented in Table S1. (C) Western blots of microneme secretion assays comparing ethanol-induced secretion of MIC2 with different concentrations of ethanol for the wild-type strain (WT), the TgDJ-1 knockout strain (ΔTgDJ-1), and the TgDJ-1 knockout strain complemented with wild-type TgDJ-1 (ΔTgDJ-1WTcomp). GRA7, loading control. (D) Quantification of induced microneme secretion assays; data represent means ± SD for results from n = 3 experiments. Significance was calculated for a two-way statistical ANOVA for comparison of the different strains (WT, ΔTgdj-1, and ΔTgdj-1WTcomp) (****, P ≤ 0.0001) and the different proportions of EtOH (0%, 0.1%, and 1%) (****, P ≤ 0.0001). Post hoc pairwise comparisons of the different strains at each ethanol concentration were performed using Tukey’s multiple-comparison test and the data presented in Table S2. (E) Western blot comparing expression levels of CDPK1 in wild-type (WT), ΔTgdj-1, and ΔTgdj-1WTcomp parasites. SAG1, loading control. (F) Quantification of CDPK1 expression levels. Data represent means ± SD for results from n = 3 experiments. Significance was calculated for an ordinary one-way statistical ANOVA to assess the difference between the means (***, P ≤ 0.001). Results of a post hoc Tukey’s multiple-comparison test for pairwise comparisons of wild-type (WT), Tgdj-1, and ΔTgdj-1WTcomp parasites are presented. ns, not significant. **, P ≤ 0.01. ***, P ≤ 0.001.