Parasites were seeded overnight (∼16–18 h) and fixed with methanol as previously described (Gubbels et al., 2006 (link)). The following antibodies were used in this study: Myc (MAb 9E10, mouse, 1:50; Santa Cruz); HA (3F10, rat; 1:3000; Roche); TgIMC3 (rat, 1:2000; Anderson-White et al., 2011 (link)); TgNuf2 and TgNdc80 (guinea pig, 1:2000; Farrell and Gubbels, 2014 (link)); SFA (rabbit, 1:1000; kindly provided by Boris Striepen, University of Georgia; Francia et al., 2012 (link)); TgNek1 (1:1000; Chen and Gubbels, 2013 (link)); HsCentrin (rabbit, 1:1000; kindly provided by Iain Cheeseman, Whitehead Institute); Ty (mouse, 1:1000; kindly provided by Sebastian Lourido); TgEB1 (rat, 1:3000; this work). Alexa Fluor A488, A568, A594, A633, and A647 secondary antibodies were used. We used 4′,6′-diamidino-2-phenylindole (DAPI) to stain nuclear material. Images were acquired using a Zeiss Axiovert 200M wide-field fluorescence microscope equipped with a Plan-Fluor 100×/1.45 NA oil objective and a Hamamatsu Orca-Flash 4.0LT camera. For superresolution structured illumination microscopy (SIM), a Zeiss Elyra S.1 microscope equipped with a Plan-Apochromat 63×/1.40 oil objective and a PCO-Tech pco.edge 4.2 sCMOS camera in the Boston College Imaging Core was used in consultation with Bret Judson. Images were acquired and processed in Zeiss ZEN v. 2.3 software using the standard mode.