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. 2006 Oct;80(19):9455–9464. doi: 10.1128/JVI.01149-06

FIG. 11.

FIG. 11.

Interaction of F9 with the entry/fusion complex. (A) BS-C-1 cells were infected with vF9-V5 or VACV WR. After 24 h, the cells were disrupted with a Dounce homogenizer and postnuclear supernatants were centrifuged at 100,000 × g for 1 h. The pellet was extracted with PBS-NP-40, and the soluble fraction was immunopurified on agarose beads conjugated to V5 antibody. Proteins were resolved by SDS-PAGE, transferred to a nitrocellulose membrane, and incubated with anti-V5 antibody conjugated to peroxidase and primary rabbit antibodies to the A16, A21, A28, and L5 proteins followed by peroxidase-conjugated secondary antibody to rabbit IgG. (B) BS-C-1 cells were infected with vA28-HAi/H2-V5 in the presence or absence of IPTG. The NP-40-PBS-soluble extract of the membrane-enriched fraction was immunopurified on agarose beads conjugated to V5 antibody and resolved by SDS-PAGE. Proteins were transferred to a nitrocellulose membrane and detected by Western blotting with peroxidase-conjugated anti-V5 or anti-HA antibody or with anti-F9 rabbit antibody followed by peroxidase-conjugated anti-rabbit IgG.

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