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P-K254R affects viral RNA transcription. P-K254R was compared to P-WT using a transcription-only minigenome system. The plasmids were transfected similarly to the normal PIV5 minigenome system. A dual-luciferase assay was carried out at 2 days posttransfection. Aliquots of the cell lysates were used for immunoblotting with anti-NP and anti-P antibodies.
