TgPuf1 expression and subcellular localization in the Pru∆Ku80 parasite. (A) Confirmation of the C-terminal HA x 3-tagging of the endogenous TgPuf1 locus. Two clones (C1 and C2) were probed with the anti-HA antibody. Lysate from wild-type Pru strain parasite was included as a HA-negative control. (B) Expression of TgPuf1 in tachyzoites and bradyzoites. The two stages of the parasite were differentiated by antibodies against BAG1, a protein expressed specifically in the bradyzoite stage. Anti-β-tubulin antibody served as a protein loading control. (C) Subcellular localization of TgPuf1 in tachyzoites and bradyzoites.