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. 2022 Jun 9;13:3208. doi: 10.1038/s41467-022-30988-z

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© The Author(s) 2022

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 — a Representative image of CSP relative expression in Plasmodium yoelii CSP_wt and CSP_mut oocysts from mosquitoes (n = 40) detected by western blot at days 7, 9, and 12 post blood meal (upper); CSP relative expression was statistically analyzed (lower). b Representative immunofluorescence image of oocyst (scale bar 10 μm) and hemolymph sporozoites (scale bar 5 μm) from P. yoelii CSP_wt or CSP_mut parasite-infected mosquitoes stained with anti-CSP-N and anti-CSP-C terminal antibodies. c Representative immunofluorescence image of in vitro nitration of hemocytes from naïve mosquitoes (n = 20) stimulated by midguts (n = 20) from P. yoelii CSP_wt or CSP_mut parasite-infected mosquitoes at day 5 PI (left). WGA was used to distinguish hemocytes, and hemocyte nitration was determined using anti-nitrotyrosine staining; scale bar 5 μm. Percentage of the nitrated hemocytes was statistically analyzed (right). d GNBP-B1, Rel1, and TEP1 expression in hemocytes from naïve mosquitoes (n = 20) following incubation with midguts (n = 20) from P. yoelii CSP_wt or CSP_mut parasite-infected mosquitoes at day 5 PI determined using real-time PCR. e Representative immunofluorescence image of the nitration of hemocytes from naïve mosquitoes (n = 20) incubated with midguts from P. yoelii CSP_mut parasite-infected mosquitoes (n = 20) at day 5 PI with or without NOX5 silenced (left); scale bar 5 μm. Hemocyte nitration was determined as described in c and statistically analyzed (right). f GNBP-B1, Rel1, and TEP1 expression in hemocytes from naïve mosquitoes (n = 20) with or without dsNOX5 incubated with midguts (n = 20) from P. yoelii CSP_mut parasite-infected mosquitoes at day 5 PI. A two-sided Student’s t-test was used for the statistical analysis. The data were presented as the means ± SD; ns, no significance; p-values were shown. The data were combined based on three independent experiments (a, c–f) or two independent experiments (b). Source data are provided as a Source Data file.