Figure 6.

The central region of ORF34 is required for interactions with ORF18, ORF31, ORF23, and ORF66, while the C-terminal domain is required to interact with ORF24. (a) Schematic representation of 2xS-tagged ORF34 deletion mutants used in the mapping experiments. The end amino acids are numbered. (b–f) 293T cells were co-transfected with expression plasmids of 2xS-ORF34 mutant and 6xMyc-ORF24 (b), 3xFLAG-ORF31 (c), 3xFLAG-ORF18 (d), or 3xFLAG-ORF66 (f). Transfected cells were lysed, and cell lysates were subjected to pull-down assays using S-protein-immobilized beads that capture the 2xS-ORF34. Obtained precipitates including 2xS-ORF34 deletion mutants were probed with indicated antibodies to detect interaction. (b–f) Original images of blotting data are shown in Supplementary Figs S10–S14.