Figure 5. AMOT p130 is required for efficient release of wild type HIV-1.
Left panels are western blots showing 293T cellular levels of endogenous ALIX (panel 1, anti-ALIX), TSG101 (panel 2, anti-TSG101), AMOT (panel 3, anti-AMOT), GAPDH (panel 4, anti-GAPDH, loading control) and levels of HIV-1 Gag proteins (panel 5, anti-MA and anti-CA). Cells were co-transfected with a control siRNA (lane 1) or with an siRNA that depleted ALIX (lane 2), TSG101 (lane 3) or AMOT p130 and p80 (lanes 4–6), with expression vectors for wild type HIV-1 (lanes 1–6) and with an siRNA-resistant expression constructs for HA-AMOT p130 proteins (with wild type HA-AMOT p130 in lane 5 and an HA-AMOT p130ΔPPXY protein with inactivating point mutations in all three PPXY motifs in lane 6). Right panels show corresponding levels of extracellular virion-associated Gag proteins (upper panel, western blot, anti-MA and anti-CA) and viral titers (lower panel), n = 4.