Eukaryotic cell lines and biG-biS viruses
Plasmids
Protein expression in eukaryotic cells
Protein complementation assays
Analysis of viral growth
Minigenome assays
Expression and purification of MD variants
Far-UV CD studies
Analytical ultracentrifugation
Crystallization and x-ray data collection of the IPKI variant
Structure determination, refinement, and analysis
Structure modeling and molecular dynamics simulations of MDs
Statistical analyses
BSR-T7 cells were plated into a 96-well plate at 2 × 104 cells per well and transfected the next day with plasmids coding for Flag/L[Renilla] and a P variant with jetPRIME (Polyplus). Luciferase signals were measured 24 hours later according to the manufacturer’s protocol (Renilla Luciferase Assay System, Promega).