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. 2011 Sep;85(18):9543–9554. doi: 10.1128/JVI.00785-11

Fig. 8.

Fig. 8.

Dimerization analysis of the WhNV B2 protein and its mutants. (A) Chemical cross-linking assays. (Left) Following a 30-min treatment with the cross-linking agent glutaraldehyde, 2 μg MBP-B2, MBP-B2ΔN20, or MBP was separated by 12% SDS-PAGE. (Right) Western blotting using anti-B2 polyclonal antibodies was conducted for further confirmation. The locations of the B2 monomer and dimer are indicated. (B) Far-Western blotting showing B2 self-interaction between two different recombinant B2 proteins. After being separated by 12% SDS-PAGE and transferred to PVDF membranes, 2 μg renatured His-B2 or His-B2ΔN20 was overlaid with 100 μg MBP-B2 or MBP as indicated above the panel and probed with anti-MBP antibody. Lane 1, MBP, a positive control to test the valence of MBP antibody used for blotting; lane 2, His-B2; lane 3, His-B2ΔN20; and lane 4, His-B2 overlaid with MBP as a negative control to eliminate the possibility of an interaction between B2 and MBP. (C) Far-Western blotting showing B2 self-interaction between His-B2 and MBP-B2 or its mutants in the absence (dsRNA −) or presence (dsRNA +) of 50 μg 500-bp dsRNA. As overlaid proteins, MBP-B2 or its mutants are indicated above the panel.