Figure 2

The involvement of Vps4A and Vps4B in HTLV-1 Gag budding. A. 293T cells were cotransfected with pK30-Gag and the expression plasmid for Vps4AEQ or Vps4BEQ, or the empty vector as a control. Extracellular VLPs were pelleted from the culture fluids. VLP-associated or cell-associated Gag was detected by western blotting (WB) using anti-HTLV-1 p19 monoclonal antibody. C. 293T cells were cotransfected with pK30-Gag and the expression vector for wild-type Vps4A or Vps4B, or the empty vector as a control. The proteins were detected as described in A. B and D. Intensities of the bands corresponding to cell- and VLP-associated Gag in A and C were quantified using the LAS3000 imaging system (Fuji film). The efficiency of Gag-induced VLP budding in cells cotransfected with pK30-Gag and control vector (VLP/Cellular) was set to 1.0. The data represent averages and standard deviations (SD) of 3 independent experiments.