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. 2020 Jul 3;3:351. doi: 10.1038/s42003-020-1081-3

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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. 5 — Protein expression analysis of PYYM_0017400.GFP knock-in cell line. a For Western blot analysis trophozoite-stage parasites were enriched by centrifugation on a Histodenz gradient. Cells were subsequently lysed using saponin resulting in a parasite (P) and supernatant (S) fraction. Exported protein is expected to be seen in the S fraction. A protein of size 63 kDa which correspond to the size of the full-length PYYM_0017400.GFP protein is detected in both supernatant and pellet fraction. The red band corresponds to EXP2 which is a parasitophorous vacoule protein used as a control for the lysis. b Live trophozoites observed under florescence microscopy showed PYYM_0017400.GFP localizing in the PVM and in punctate dots. PYYM_0017400.GFP is found to surround individual merozoites in late schizonts and free merozoites. DNA was stained using Hoechst33342.