Fig. 4

The Gag–RPL7 interaction is independent on Gag myristoylation and oligomerization. Lysates from HeLa cells transfected with pcDNA (lanes 2, 7) or plasmids coding for Gag (lanes 3, 8 and 11), Gag-G2A (lanes 4, 9) and Gag-M369A (lanes 5, 10). Control: lysate from non-transfected HeLa cells (lanes 1, 6). The lysates were subjected to IP (1 mg of total protein) using anti-p24 antibody followed by western blot where 20 µg of total protein (input) and resuspended beads were analyzed with anti-p24, anti-RPL7 and anti-GAPDH antibodies revealed by protein A-HRP, anti-rabbit HRP conjugate or anti-mouse HRP conjugate, respectively. All Gag proteins (WT or mutants) interact with endogenous RPL7 (lanes 8–10). Neither Gag nor RPL7 bands were observed in non-transfected cells (lane 6), in pcDNA transfected cells (lane 7), or in the control of beads without anti-p24 (lane 11). X Heavy chain of anti-p24 used to IP Gag and Gag derivatives