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. 2023 Sep 28;14:6078. doi: 10.1038/s41467-023-40948-w

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Schematic of the ROCY1 mRNA indicating the location of the putative NLS (blue bar) and CCCH motifs (yellow bars). Shown below are AlphaFold2 predictions for the 3 CCCH motifs, with each cysteine and histidine shown with side chains. N- and C-terminal directionality is indicated as are the locations of each CCCH motif. b Quantification of the percentage of DBA-positive cysts among HA-positive vacuoles expressing the WT pGRA NHA-ROCY1 (N = 104) or the CCCH mutant form of pGRA NHA-ROCY1 (N = 107) under control (pH 7.2) conditions. The CCCH mutations ablated the ability of pGRA NHA-ROCY1 to induce cyst formation in the ROCY1 knockout background. c Mean and standard deviation of background-normalized HA-derived fluorescence intensity of TgPRUΔROCY1 expressing either WT pGRA NHA-ROCY1 (N = 24) or the CCCH mutant form of pGRA NHA-ROCY1 (N = 19) showing similar staining intensity (Student’s T-test P = 0.2). d Representative images of vacuoles containing PRUΔROCY1 parasites expressing the wild type or CCCHmutant form of the pGra_NHA_ROCY1 plasmid construct. Scale bar: 10 μm (same for all images). This experiment was performed once with this genetic background. Data are provided as a Source Data File.