Fig 6.

The YFV RC is on a detergent-insoluble fraction. (A) Schematic of experimental procedures for panels B and C. WB, Western blotting; IFA, immunofluorescent staining. (B) Proteinase K digestion of YFV-infected cells. SW13 cells were infected with YFV (MOI, 5) and permeabilized in situ 1 day later with 50 μg/ml digitonin or solubilized with 1% Triton X-100. The supernatant (S) was collected for Western blotting. The remaining detergent-insoluble cell material (IS) was then washed, scraped, and treated with 10 μg/ml proteinase K or not treated. Proteins were TCA precipitated and analyzed by Western blotting with the antibodies indicated. The values to the left are molecular sizes in kilodaltons. (C) After in situ permeabilization and washing, cells were fixed and permeabilized with 0.5% Triton for immunofluorescence analyses and examination by confocal microscopy. The results shown are representative of multiple similar experiments. Bar, 10 μm.