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. 2020 Sep 23;11:4813. doi: 10.1038/s41467-020-18624-0

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© The Author(s) 2020

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, b The DegP2 coding sequence was replaced with YFP to generate the ∆DegP2 line, which was subsequently complemented with an HA-tagged copy of DegP2 (dark green) under the regulation of the TUB1 promoter (a). Separately, the endogenous locus of DegP2 was first tagged with the Ty epitope, and a point mutation was introduced at the catalytic serine (b). DegP2-HA, DegP2-Ty, and DegP2^S569A-Ty co-localized with the mitochondrial marker mtHSP70. Merged image additionally displays YFP (green) for ∆DegP2 strains and DNA stain (blue) for the HA-stained samples. Scale bar is 5 μm. c A DegP2-inducible mutant constructed using the U1 system. Staining for the HA tag appended to the DegP2 cKD locus showed the expected localization of the protein to the mitochondrion. Individual parasites are visualized by staining for CDPK1. Scale bar is 5 μm. d Immunoblotting for DegP2’s HA tag showed robust depletion of DegP2 48 h after a 2 h rapamycin treatment. CDPK1 was used as a loading control.