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. 2019 Jul 30;10:3396. doi: 10.1038/s41467-019-11388-2

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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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Fig. 2 — ANP32A variants differentially impact avian-signature IAV replication and selection. a Genotype of the A549-ANP32A_KO cell-clone used in this study. The target site of the crRNA is depicted. Four distinct ANP32A alleles were detected in A549s suggesting duplication of chromosome 15. b Western blot analysis of parental A549 (Ctrl) and A549-ANP32A_KO (KO) cells for ANP32A, ANP32B, and actin. c Western blot analysis of A549-ANP32A_KO cells stably reconstituted with the indicated FLAG-ANP32A constructs or empty vector. d–f Viral growth kinetics of rWSN-based viruses expressing PB2-627E or PB2-627K in the indicated ANP32A-reconstituted cell-lines (MOI = 0.001 PFU/cell). g–i Competition assays between rWSN-based viruses expressing PB2-627E or PB2-627K (5:1 input ratio) in the indicated ANP32A-reconstituted cell-lines. P0 represents input. Percentage of PB2-627K/E was determined by NGS. In panels d–i, mean values from three independent experiments are plotted with lines, and the individual data points are shown. For panels b and c, representative data from two independent experiments are shown. Source data for panels b–i are provided in the Source Data file