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. 2017 Jul 20;7:6026. doi: 10.1038/s41598-017-05917-6

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© The Author(s) 2017

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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Midgut invasion assays of Δpbp47 parasites. (A) Fluorescence microscopy of ANKA 507m6cl1 (wt) and Δpbp47 ookinetes stained with P28 antibody in the A. gambiae midgut epithelium 24–26 hours post blood feeding. Images are taken at 100x magnification, and inset images at 400x magnification. (B) Light microscopy images (100x magnification) of melanized ANKA 507m6cl1 (wt) and Δpbp47 parasites in CTL4 silenced A. gambiae mosquito midguts. (C) Left: load of melanized ANKA 507m6cl1 (wt) and Δpbp47 ookinetes in the midguts of CTL4 silenced mosquitoes (left). Red lines show the median ookinete load. Right: forest plots of melanized ookinete loads as determined by GLMM analysis. The variation of the fixed effect estimate in each (squares) and all (diamonds) replicates (R) is shown ( ± 95% confidence interval, glmmADMB). The square size is proportional to the sum of midguts analyzed in each replicate. (D) Confocal images (630x magnification) of a Δpbp47 melanized ookinete in the midgut of CTL4 silenced A. gambiae, stained with the P28 antibody (red) and DAPI (blue). BF is bright field. Scale bar is 5 µm.