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. 2022 Sep 30;12:1011692. doi: 10.3389/fcimb.2022.1011692

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Copyright © 2022 De Meulenaere, Prajapati, Villasis, Cuypers, Kattenberg, Kasian, Laman, Robinson, Gamboa, Laukens and Rosanas-Urgell

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Schematic representation of the experimental approach. P. vivax–infected red blood cells are cultured until the majority have matured into schizonts, after which schizont stages are enriched and used for invasion assays; if enough schizonts are available, they are stored for subsequent mRNA sequencing. Three different invasion assay setups are used: invasion into SAO reticulocyte-enriched RBCs (reRBCs), band 3 blocking with a polyclonal antibody (pAb), and band 3 blocking with a PvTRAg38 peptide. The number of newly invaded ring- and young trophozoite-stage parasites is counted by light microscopy, and compared to the paired control (non-SAO reRBCs without antibody/peptide), resulting in a normalized percentage of invasion inhibition. Figure created with BioRender.com.