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. 2019 Dec 4;9:18300. doi: 10.1038/s41598-019-54613-0

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© The Author(s) 2019

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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Constitutive expression of Plasmodium berghei PDI-Trans, and localization on the surface of gametocytes and ookinetes. (A) RT-PCR analysis of PDI-Trans in asexual blood stages using the non-gametocyte producing strain 2.33; non-activated (Gc-) and activated (Gc+) gametocytes; purified in vitro ookinetes and day 21 salivary gland dissected sporozoites. The analysis was complemented with alpha-tubulin loading controls (B). PCR confirmation of integration of egfp into the PDI-Trans locus. Oligonucleotides 35 and 14 were used to detect integration. Oligonucleotide 91 and 92 were used to detect DHFR presence, pbs25 oligonucleotides were used as positive controls. P. berghei WT 2.34 gDNA was used as a negative control for integration. (C) IFA of fixed, non-permeablised PDI-Trans-GFP parasites probed with anti-GFP; exflagellating male gametocytes (top) and ookinetes (bottom). Each panel shows an overlay of GFP fluorescence (green) and DNA labelled with DAPI (blue). White scale bar = 5 μm.