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. 2024 Jan 26;15:793. doi: 10.1038/s41467-024-44967-z

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a AP2XII-1-mAID-6HA localizes to the nucleus in tachyzoites (32 h post-infection) and bradyzoites (3 days post-infection) and is efficiently depleted when the parasites are treated with IAA. Magenta, anti-IMC1; green, FITC-DBL; red, anti-HA. Scale bar, 10 µm. b Quantification of bradyzoite differentiation in the indicated strains following 3 days of exposure to an alkaline culture medium. Data are presented as the mean ± SD from at least three independent biological replicates and the percentage of DBL-positive vacuoles was calculated based on a minimum of 150 vacuoles per replicate. The percentage of vacuoles scoring “DBL-high” was used for comparison and analyzed by two-tailed, unpaired t test, *p = 0.0379 (AP2XII-1-mAID, -IAA vs +IAA), **p = 0.0020 (209500-mAID, -IAA vs +IAA), ***p = 0.0003 (214140-mAID, -IAA vs +IAA), *p = 0.0449 (275680-mAID, -IAA vs +IAA), **p = 0.0022 (TIR1 -IAA vs 275680-mAID -IAA), ****p < 0.0001 (TIR1 -IAA vs AP2XII-1-mAID -IAA). c Representative images of the plaques formed by the indicated parasites grown in HFF monolayers for 8 days. d Relative size of the plaques detected in c. Data represents the mean ± SD from three independent experiments. Statistical significance was tested by two-tailed, unpaired t test, **p = 0.0019 (TIR1 -IAA vs AP2XII-1-mAID -IAA), **p = 0.0074 (209500-mAID -IAA vs +IAA), **p = 0.0014 (214140-mAID -IAA vs +IAA), **p = 0.0067 (TIR1 -IAA vs 275680-mAID -IAA), ****p < 0.0001. e The subcellular location of the indicated proteins in the mature merozoites. The indicated parasites were allowed to infect HFFs for 4 h followed by incubation in an alkaline medium without CO₂ for 3 days with IAA. Green, anti-IMC1; red, anti-Ty; Scale bar, 10 µm. f Genome-wide analysis reveals that AP2XI-2, AP2XII-1 and MORC associate with the transcriptional start sites (TSS) of the genes. The indicated parasites treated with or without IAA for 48 h were used to assess the genomic loci of AP2XI-2, AP2XII-1 or MORC. g Integrated genome browser view of AP2XI-2, AP2XII-1 and MORC enrichment across T. gondii chromosome Ia. The data of MORC-HA Rep1 and MORC-HA Rep2 were obtained from the GEO under accession number GSE136060¹⁴. Source data are provided as a Source data file.