Uninfected or infected Vero cells (producer cells, MOI 1 PFU/cell at 24 h) in 24-well plates were transfected with 100 ng of DVG replicon RNA, WT replicon (positive control), or inactive replicon (negative control), using TransIT-mRNA transfection reagent (Mirus Bio). 48 h p.t. the supernatant was endonuclease-treated (25 Units/μl BaseMuncher, Expedeon) and used to infect naïve Vero cells in 24-well plates (recipient cells). Replicon activity in recipient cells was assessed 24 h p.i. Donor and recipient cells were lysed in passive lysis buffer (Promega) and luciferase activity measured using the Nano-Glo luciferase assay system (Promega) in a Tecan infinite M200 pro plate reader (Tecan group).

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