Figure 5

HA-endophilin 2 is incorporated inside Mo-MuLV virions. Equal amounts of purified virion particles were subjected to subtilisin digestion at 0, 1, 10 or 100 μg/ml. Protease inhibitors PMSF and aprotinin were subsequently added to terminate the digestion. (a) Virion particles after digestion overnight at room temperature were sedimented through a 25% sucrose cushion, and the proteins in the pellets were analyzed by western blotting with anti-HA, anti-capsid and anti-p15E envelope antibodies. (b) Virion particles were subject to subtilisin treatment in the presence of 0.2% NP-40, and then were directly analyzed by western blotting with anti-HA and anti-capsid antibodies.