Fig 1: ANP32A variants differentially impact avian-signature IAV replication and selection. a Genotype of the A549-ANP32AKO 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-ANP32AKO (KO) cells for ANP32A, ANP32B, and actin. c Western blot analysis of A549-ANP32AKO 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