Figure 2

Phylogenetic analysis, gene tagging identification and intracellular localization of TgOTU7. A Phylogenetic tree of OTU7 of different Apicomplexa species. A phylogenetic tree was constructed based on the amino acid sequences with the neighbour-joining algorithm using MEGA X. Bootstrap analysis was performed with 1000 replicates. The red star indicates OTU7 of T. gondii. The scale bar indicates the evolutionary distance between the sequences. B PCR analysis of 6 × HA-tag insertions at the 3′ end of the TgOTU7 gene. C Western blot identification of the TgOTU7-6HA protein using rabbit anti-HA antibodies. The expected size of the TgOTU7-6HA proteins is highlighted by a red star. The bands of smaller size may be due to processed or degraded forms of TgOTU7-6HA. D Immunofluorescence assay revealing the intracellular localization of endogenously tagged TgOTU7. Parasites were stained with rabbit anti-HA (green), mouse anti-IMC1 (red) and DAPI (blue).