The expression of P protein constructs was verified by Western blot using a polyclonal rabbit MeV P protein as detailed (9). The ability of P and domains of P to associate with N protein in cellula was tested by the Gaussia-based protein complementation assay, as detailed elsewhere (8, 9, 20). Briefly, plasmids encoding P protein fused to either the C terminus or N terminus of the N-terminal glu1 domain, and N protein fused to the C terminus of glu2 domain by polymerase chain reaction and In-Fusion (Clontech) recombination. Glu1 and glu2 fusion constructs were transfected in human embryonic kidney–293T cells (from CelluloNet BioBank BB-0033-00072, SFR BioSciences), and Gaussia luciferase activity was measured after 2 days of culture. The results were expressed in normalized luminescence ratio as described (58). Each test was carried out three times with each condition applied in triplicate.

Minigenome assay was performed in BSR-T7 cells, which constitutively express the T7 polymerase (59), as detailed previously (9). The minigenome included the leader and trailer sequences separated by a first gene encoding the firefly luciferase and a second gene encoding the Gaussia luciferase, the expression of which relies on the edition of the gene transcript. The two genes are separated by an intergenic junction elongated by 324 nucleotides (9). Briefly, plasmids encoding for MeV minigenome RNA of (+) or (−) polarity, N, L, and P protein under the control of T7 promoter were transfected together in BSR-T7 cells. Two days later, the expression of the two-luciferase reporter gene was recorded by luminescence measurement in the presence of their specific substrate. Each test was performed three times with each condition performed in triplicate.

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