Fig. 2: In vitro characterization of viruses carrying the P136S, A156T or P136S/A156T substitutions. | npj Viruses

Fig. 2: In vitro characterization of viruses carrying the P136S, A156T or P136S/A156T substitutions.

From: Impact of naturally occurring hemagglutinin substitutions on antigenicity and fitness of influenza A(H5N1) virus

Fig. 2

a Replication kinetics of rTX37 and rTX37-HA-P136S/A156T in NhNE cells at 37 °C and 32 °C. b Replication kinetics of rTX37-PB2-627E and rTX37-PB2-627E-HA-P136S/A156T in NhNE cells at 37 °C and 32 °C. c, d Competition assays in NhNE cells at 32 °C. Virus inocula were prepared by mixing two viruses at 1000 TCID₅₀ with ratios of 1:1 (c) and 1:3 (d) and used to infect cells in four or six technical replicates. Apical washes were collected daily, and RNA extracted from Day 1 and Day 4 samples was analyzed by next-generation sequencing (NGS). Pie charts show the mean frequencies of HA variants in apical wash samples: wild type (red), P136S/A156T (blue), P136S (pink) and A156T (sage). The average percentage of wild-type virus is shown in the figure. Statistical analysis of wild-type virus frequencies between the inoculum and samples collected on days 1 and day 4 was performed using one-way ANOVA followed by Dunnett’s multiple comparisons test. Error bars indicate SD. “*” indicates significant difference (P < 0.05). “**” indicates very significant difference (P < 0.01).

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