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. 2023 Feb 21;6:205. doi: 10.1038/s42003-023-04566-y

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© The Author(s) 2023

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. 3 — a, b Screen of 15 non-essential P. berghei kinases in loss-of-function mutants using the ex vivo maturation assay. a Percentage of mature segmented schizonts and (b) merozoite numbers per schizont in wild-type and kinase knockout lines (Δnek4: PBANKA_061670; ΔeIK1: PBANKA_1308400; ΔeIK2: PBANKA_0205800) cultured in MetS vs MetD media. c, d Quantification of (c) P. berghei intra-erythrocytic developmental stages and (d) merozoite numbers of P. berghei wild-type and kinase mutant ΔeIK2, ΔeIK1 and Δnek4 parasites cultured in MetS or MetD media. e, f Percentage of mature segmented schizonts and (f) merozoite numbers in wild-type and kinase knockout parasite lines cultured in IleS vs IleD media. g, h Quantification of (g) P. berghei intra-erythrocytic developmental stages and (h) merozoite numbers of P. berghei wild-type and kinase mutant ΔeIK2, ΔeIK1 and Δnek4 parasites cultured in IleS or IleD media. a, b, e, f Kinase knockout mutants were analyzed in triplicate and averaged, with each data point representing the a, e mean percentage of mature schizonts or b, f the mean merozoite number produced per schizont. Error bars represent the SE Diff or SD between replicates, respectively. n = 3 independent experiments for wild-type, ΔeIK2, ΔeIK1 and Δnek4 or n = 2 for the other knockout lines. c, g Parasite staging was performed in triplicate, averaged and repeated in c. n = 3 or g. n = 2 independent experiments. Data is shown as mean percentage of parasites at each developmental stage, with error bars representing SEM. Individual data points are represented in Supplementary Fig. 9a, b. d, h Data is represented as box-whisker plot (10–90 percentile) of mean merozoite number per schizont ±SD (2-way ANOVA). Boxplots show the data of d n = 3 or h n = 2 independent experiments and for the following number of schizonts (N), wild-type: MetS, N = 148; MetD, N = 148; IleS, N = 114; IleD, N = 133; ΔeIK2: MetS, N = 85; MetD, N = 158; IleS, N = 70; IleD, N = 68; ΔeIK1: MetS, N = 220; MetD, N = 238; IleS, N = 75; IleD, N = 94 and Δnek4: MetS, N = 106; MetD, N = 106; IleS, N = 90; IleD, N = 102.