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. 2024 May 23;15:4385. doi: 10.1038/s41467-024-48685-4

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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. 6 — a Stabilization of BFD1 or BFD2 in WT or ∆eif1.2 parasites. b Western blot analysis of lysates from parasites treated with vehicle control (100% ethanol) or 3 µM Shield−1 for 4 days. Actin, loading control. The actin and DD-BFD1-Ty were from the same sample but run on separate blots due to a bubble in one of the original actin bands on the same blot as DD-BFD1-Ty. Uncropped images for both the current actin and previous actin blots are provided in the Source Data for (b). c–f Quantification of western blot results. AU, arbitrary units. Data represent the mean ± s.d. n = 3 biological replicates. The DD-BFD1-Ty/Actin ratio was analyzed using a linear regression model. The DD-HA-BFD2/Actin or BAG1/Actin ratios were analyzed using a linear mixed model. NS, not significant (P > 0.05). e The BAG1 signals were captured when BAG1 bands in all lanes in the western blots were not saturated. Here, BAG1 levels in Shield−1 treated DD-BFD1-Ty∆eif1.2 were notably low and showed no significant difference compared to the vehicle-treated group. f With longer exposure of the western blots to ensure BAG1 signals for DD-BFD1-Ty∆eif1.2 parasites not saturated, Shield−1 treated DD-BFD1-Ty∆eif1.2 exhibited significantly higher BAG1 levels compared to the vehicle treated control. g Representative vacuoles of parasites treated with vehicle or 3 µM Shield−1 for 4 days. Bar, 10 µm. h, i Measurement of mean fluorescence intensity for DBA and BAG1 within individual vacuoles. Mean ± s.d. plotted for 3 biological replicates. A minimum of 100 vacuoles were quantified for each condition within each replicate. Data were analyzed with a linear mixed model. j Model for the role of eIF1.2 in T. gondii differentiation. Upon stress, eIF1.2 enhances the expression of key bradyzoite-specific factors, such as BFD1 and BFD2, to drive T. gondii differentiation. Source data are provided as a Source Data file.