Fig 4.

Far-Western blotting to detect interacting recombinant T4SS proteins with specific targets. The indicated purified recombinant T4SS proteins (1 μg) were separated on 4 to 20% gradient SDS-PAGE gels, transferred to a nitrocellulose membrane, and incubated with 10 μg of target protein rVirB9-1 (A), rVirB9-2 (B), and rVirB10 (C). The interactions were detected with E. coli-adsorbed rabbit antisera (1:500) specific for VirB9-1, VirB9-2, and VirB10. Blots were developed with goat anti-rabbit IgG(H+L). Scanned images were rearranged and presented as shown. In panel A, the results for VirB11 and VirD4 were obtained from a separate experiment. An asterisk indicates an interacting protein partner to the target protein with the following predicted molecular masses: rVirB4-1 (90 kDa), rVirB4-2 (91 kDa), rVirB6-1 F1 (90 kDa), rVirB8-2 (23 kDa), rVirB9-1 (46 kDa), rVirB9-2(46 kDa), rVirB10 (65 kDa), rVirB11 (38 kDa), rVirD4 (93 kDa), and MSP5 (28 kDa).