Figure 5: Bat NP-specific amino acids in SC35M NP cause impaired packaging.

(a,b) Packaging efficiency of a single minigenome supported by SC35M NP (NP), CH2, NP7 or NP7-R31G in a VLP-based packaging assay. Reconstitutions of SC35M VLPs were carried out in the presence a GFP-encoding reporter segment flanked by IS of the NP segment (IS−GFP) or in addition with the bundling signals (BS) in the 5′ and 3′ NP ORF (IS+BS−GFP). After infection of MDCKII cells and subsequent superinfection with wt SC35M, GFP-positive cells were quantified by FACS. NS, not significant; *P<0.05; **P<0.01. (c,d) Packaging efficiency mediated by SC35M NP (NP), NP7, CH2 or NP7-R31G in the presence of the reporter segment IS+GFP (R) (c) or IS+BS−GFP (R) (d) and, if indicated, in the presence of the remaining seven wild-type genome segments (+7). NS, not significant; *P<0.05; **P<0.01; ****P<0.0001. Student’s t test was used for two-group comparisons. Error bars indicate the mean and s.d. of at least three independent experiments.