Fig 3. The 2716-Ymg-03 strain replicates in HS-deficient and hSCARB2-overexpressing cells without attenuation or mutation in VP1-145.

The 2716-Ymg-03 strain was passaged in genetically modified RD-A cells. (A–E) To detect SNVs in passaged virus populations, NGS reads were mapped to the consensus sequence of the p-0 virus and analyzed using LoFreq and SnpEff software. The abundance of each annotated SNV was plotted on the corresponding position in the virus genome. Vertical dotted lines indicate borders separating genetic regions (5’UTR, P1, P2, P3, and 3’UTR). Horizontal dotted lines indicate the 1.5 IQR of the abundance of each SNV mapped to the graph. Arrowheads indicate the position corresponding to VP1-145. (F–J) The abundance of VP1-145 mutants in the virus population before and after passage, estimated from NGS data using ShoRAH software. (K–O) The passaged 2716-Ymg-03 strains were used to infect hSCARB2 tg mice (10⁶ TCID₅₀, ip). Paralysis and death of the infected mice were observed over 14 days.