Extended Data Figure 8 : Mutant A/WSN/33 virus rescue experiments.

From: Structural basis of an essential interaction between influenza polymerase and Pol II CTD

Extended Data Figure 8

The titers and plaque phenotypes of the recombinant A/WSN/33 viruses were determined on reverse genetics supernatants (left) or upon a single plaque purification followed by viral amplification (right), using a standard plaque assay on MDCK cells. Crystal violet staining of cell monolayers infected with the indicated viral dilutions is shown. WT, control performed with wild-type PA. For each recombinant virus, two independent viral stocks (denoted as P1 and P1′) derived from the same purified plaque were subjected to vRNA extraction, RT–PCR of the eight genomic segments and next-generation sequencing. The non-synonymous mutations detected in >30% of the reads upon alignment with the reference sequences for the wild-type virus are indicated. *Number of purified plaques leading to successful viral amplification per total number tested. †P1′ did not yield any detectable infectious virus for the R454A mutant. ‡P1 (for R454A and wild type) and P1′ (for K289A, K635A and R638A) were assayed in parallel.