Fig 1. NGS-based frequency analysis of DelVG across all segments over time in various lineages.

(A) Particles to infectivity ratios were calculated at each passage of both lineages (L1 & L2). The particles of each progeny were measured by the hemagglutination assay and divided by the infectivity, measured by TCID50. (B) Comparison of DelVG junction count (left panel) and DelVG NGS read frequencies (right panel) at each passage of all segments, both metrics were normalized to 106 mapped reads relative to the total aligned reads of each segment. (C) Heatmap showing the proportional abundance (%) of PB2-DelVGs, quantified from NGS reads, across six additional lineages together with L1 and L2. Only DelVGs reaching a relative abundance of ≥2% were analyzed. Each row represents a distinct junction and displays its abundance across the eight lineages. To facilitate interpretation, theoretical maximum and minimum values are shown at the top and bottom, respectively. White cells indicate absence of the junction in that replicate. (D) The WT/DelVG ratio was calculated (for PB2, PB1, PA segments) based on the NGS read counts and DelVG Shannon diversity was calculated and superimposed.