Construction of the TgPP6 subunits knock-out and complemented strain.

(A) Schematic illustration shows the targeted deletion of the coding region of TgPP6. (B) Diagnostic PCR3 and PCR5 confirmed the correct 5’ and 3’ region integration of homologous fragments, respectively, and PCR4 confirmed the deletion of TgPP6 coding region. (C) Complementation strategy of TgPP6C and TgPP6R into UPRT locus and isolation of clones by chloramphenicol. (D) and (E) The invasion efficiency of RHΔpp6c and RHΔpp6r strains was slightly lower than that of WT, RHΔpp6c-cp and RHΔpp6r-cp strains, while the egress rate of RHΔpp6c and RHΔpp6r strains was similar to the other strains. Data represent mean ± SD of three independent experiments and significance was analyzed by unpaired t-test. *P = 0.0396 (RHΔpp6c vs. WT); *P = 0.0448 (RHΔpp6r vs. WT); n.s., not significant.

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