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. 2024 Apr 8;15:2999. doi: 10.1038/s41467-024-47097-8

Fig. 4. TKT is essential for the lytic cycle of parasites.

Fig. 4

a Engineering of TKT strains. b Immunostaining of TKT in the DiCreTgTKT-Ty strain. Parasites were stained with anti-Ty and anti-TgALD antibodies. Scale bars = 5 μm. c Immunofluorescence staining of DiCreTgTKT-Ty and DiCre-YFP+ with anti-Ty antibody. Scale bars = 5 μm. d Expression of TgTKT-Ty in TgTKT-TyΔtkt strain. TgTKT-TyΔtkt parasites were cultured without or with rapamycin for 4 days and then stained with anti-Ty antibody. Scale bars = 5 μm. e Expression of TKT in the Δtkt mutant. The parental DiCre strain was stained with mouse anti-TgTKT and rabbit anti-TgALD, whereas the Δtkt strain (YFP positive) was stained with mouse anti-TgTKT. Scale bars = 5 μm. f-h Plaque and replication assays with indicated strains. The TgTKT-TyΔtkt tachyzoites were treated with rapamycin for 4 d to deplete TKT before analysis. n = 3 experiments, means ± SEM; unpaired two-tailed Student’s t-test (****, p < 0.0001). Intracellular replication assay (24 h infection; n = 3 assays, means ± SEM; ****, p < 0.0001, two‐way ANOVA). i Invasion efficiency of the DiCre and Δtkt strains, as determined by two-color staining to distinguish invaded vs non-invaded parasites (n = 4 experiments, each with two technical replicates, means ± SEM; ****p < 0.0001, unpaired two-tailed Student’s t-test). Source data are provided as a Source data file.