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. 2021 Apr 21;6(2):e00182-21. doi: 10.1128/mSphere.00182-21

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Copyright © 2021 Guevara et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 4 — GRA12-related proteins colocalize with GRA2 in the cyst matrix and at the cyst wall. HFF cells were infected with complemented GRA12-related HA-tagged protein strains, and in vitro cysts were differentiated for 6 h (A), 1 day (B), 2 days (C), 3 days (D), 7 days (E), or 10 days (F). Cysts containing GFP⁺ bradyzoites were located using DIC microscopy and imaged by confocal microscopy. DAPI stained host and parasite nuclei. Cysts were stained with anti-GRA2 and anti-HA antibody. Panels show DAPI and GFP, anti-GRA2, anti-HA, anti-GRA2 and anti-HA, and DIC (cyst periphery/wall indicated by white arrowhead). GRA2 colocalized with GRA12-related proteins in 100% of cysts evaluated at 6 h (n = 11 to 13), with GRA12A and GRA12B in 100% of cysts evaluated at 1 day (n = 13 to 16) and 2 days (n = 6 to 12), and with GRA12D in 100% of cysts evaluated at 3 days (n = 17). Occurrence of GRA2 colocalization phenotype was 100% in all cysts analyzed, 1 day (n = 9 to 16), 2 days (n = 7 to 12), 3 days (n = 17 to 22), 7 days (n = 10 to 24), and 10 days (n = 12 to 15). Bars = 5 μm.