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. 2014 Aug;88(16):9287–9296. doi: 10.1128/JVI.00935-14

FIG 6.

FIG 6

Protein V interacts with the membrane lytic protein VI. A Strep-tag pulldown assay was performed with 100 μg of recombinant protein V following incubation with 100 μg of recombinant protein VI or the Ad fiber knob (control). The proteins were added to Tactin resin, the resin was washed, and bound proteins were eluted with α-desthiobiotin. All of the fractions were analyzed by 4 to 20% SDS-PAGE and stained with Simply Blue. (A) SDS-PAGE of protein V-VI pulldown assay. Lanes: 1, protein V; 2, protein VI; 3, input; 4, flowthrough; 5 to 7, washes; 8 to 10, elutions; 11, molecular mass markers. (B) SDS-PAGE of protein V-fiber knob pulldown assay. Lanes: 1, protein V; 2, fiber knob; 3, input; 4, flowthrough; 5 to 7, washes; 8 to 10, elutions; 11, molecular mass markers. (C) Protein VI competition for protein V-DNA interaction. EMSA was performed with 0.5 μg of DNA and 2.5 μg of recombinant protein V and incubated with increased amounts of recombinant protein VI at 25°C for 20 min prior to analysis on a 0.8% agarose gel and then stained with SYBR green to visualize the DNA.