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Figure 3
Effects of vPIP mutations on the regulation of virus lytic replication. (a) Trans-complementation of vPIP-S virus replication by vPIP mutants. Vero cells were co-transfected with FLAG-tagged vPIP mutants in the presence of the vPIP-S virus BAC DNA (pMHV-68/vPIP-S). MHV-68 BAC DNA (pMHV-68) and vPIP-S-MR were used as a control. After 5 d of transfection, the cell lysates were subjected to Western blotting. The band intensity of the ORF45 protein was measured and normalized with that of α-tubulin using the ImageJ program. (b, c) The effect of vPIP mutants on RTA-mediated transactivation. HEK293T cells were transfected with the reporter construct containing RTA promoter (Rp-LUC) (b) or M3 promoter (M3p-LUC) (c), and vPIP mutants in the presence of RTA-expressing plasmid. Each transfection was performed in triplicate, with a β-galactosidase-expressing plasmid included as an internal control. Statistical analysis was performed using a two-sided Student's t-test (*** denotes a P value of <0.005 against WT vPIP expressing samples).

IUCrJ
Volume 5| Part 6| November 2018| Pages 866-879
ISSN: 2052-2525
https://doi.org/10.1107/S2052252518013854