Fig 7. Gag Zip fails to associate with viral-RNA-containing granules, but is rescued by a heterologous viral-RNA-binding domain.

(A) COS-1 cells transfected with indicated constructs (Set II constructs in Fig 1A) were harvested following PuroHS treatment, and the number of unspliced viral RNA copies per 1000 cells was determined for cell lysates. (B) Lysates from A were analyzed by velocity sedimentation, and the number of unspliced HIV-1 RNA copies per 1000 cells was determined for each gradient fraction and normalized to inputs in A. (C) Gradient fractions from B were subjected to IP with αGFP, and the number of unspliced viral RNA copies per 1000 cells was determined was determined for IP eluates from each gradient fraction and normalized to input in A. (D) COS-1 cells transfected with indicated constructs (Set II constructs in Fig 1A) were harvested following PuroHS treatment, and the number of unspliced viral RNA copies per 1000 cells was determined for cell lysates. (E) Lysates from D were analyzed by velocity sedimentation, and the number of unspliced viral RNA copies per 1000 cells was determined for each gradient fraction and normalized to inputs in A. (F) Gradient fractions from E were subjected to IP with αGFP, and the number of unspliced viral RNA copies per 1000 cells was determined for IP eluates from each gradient fraction and normalized to inputs in A. Positions of kD markers are shown to the right of blots. Brackets at top show S value markers, and dotted lines demarcate assembly intermediates based on their migrations in the corresponding Gag WB. Error bars show SEM from duplicate samples. Data in each column are from a single experiment that is representative of two independent replicate experiments.