Fig. 5 | Nature Communications

Fig. 5

From: Nucleotide resolution mapping of influenza A virus nucleoprotein-RNA interactions reveals RNA features required for replication

Fig. 5

vRNA regions required for PR8 replication are required for replication of contemporary avian and human IAV. a Phylogenetic analysis of segment 5. WT-PR8, WT-H7N3, or WT-pH1N1 IAV are indicated by corresponding, labeled arrows. Phylogenies were created from randomly sampled full-length segment 5 sequences downloaded from NCBI IVR. Alignment performed in MEGA (version 7) using MUSCLE. Phylogenetic trees created using the Maximum Parsimony method included in MEGA. Phylogenies were visualized in FigTree and manually annotated. Green shading represents segment sequences derived from avian viruses; red represents human viruses; and blue represents swine viruses. b Phylogenetic analysis of segment 2 performed as in a. c Relative focus area of WT-H7N3 and mutant viruses + s.e.m. The results are the average of 3 independent experiments and >60 foci per virus. Statistical significance was determined by one-way ANOVA with multiple comparisons correction (Kruskal–Wallis test). ****P < 0.001. d Viral titer (TCID50 ml−1) of 4 HA-units of WT-H7N3 and mutant virus. Results are the average of three viral titrations and two HA assays per infectious virus titration (TCID50 assay and HA titration experiments, mean + s.e.m.). Statistical significance was determined by unpaired t test. ****P < 0.001. e Relative focus area of WT-pH1N1 and mutant viruses + s.e.m. The results are the average of 3 independent experiments and >60 foci per virus. Statistical significance was determined by one-way ANOVA with multiple comparison corrections (Kruskal–Wallis test). ****P < 0.001. f Viral titer (TCID50 ml−1) of 4 HA-units of WT and mutant virus. Results are the average of three viral titrations and two HA assays per infectious virus titration (TCID50 assay and HA titration experiments, mean + s.e.m.). Statistical significance was determined by unpaired t test. **** P < 0.001

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