Figure 4

(A) Particle release from MDMs was assessed over time using a p24 antigen ELISA. The efficiency of release was plotted as percentage of extracellular p24/total p24 from 3 experiments. The majority of p24 was retained in macrophages infected with either NL4-3 or NLUdel, and total amounts of p24 (cell + sup) were similar. Right-sided graph represents one of the experiments, presented as released p24 antigen. Error bars represent SD. (B) Western blot of tetherin and actin over time following infection of MDMs. (C) Western blot of tetherin and actin over time from MDMs infected with HIV-1 BaL. MDMs were infected with HIV-1 BaL and cell lysates were harvested at the indicated time points. Western blots were performed as mentioned above. Results from 2 donors were shown as representatives of 5 independent experiments. (D) Cell surface levels of tetherin following infection with NL4-3 (dashed line) or NLUdel (solid line). Cells were stained with anti-p24 antibody to allow gating on the infected population. Results of individual experiments in HeLa (left) and MDMs (right) are shown. Below are %MFI with error bars representing SD at 48 hours postinfection (HeLa) or 4 days post-infection (MDMs), with results from three independent experiments plotted. Statistical comparison between the groups were performed by unpaired t-test using GraphPad Prism 5. See also Figure S4.