Figure 6

The inhibition of aPKC significantly decreases single-round HIV-1 infection. (A) Schematic representation of the experimental system. Briefly, 293T cells were either mock treated, or treated with aPKC inhibitor. After 4 hours, these cells were co-transfected with pNL4-3Δenv-luc or pNL4-3ΔenvΔVpr-luc and with pVSV-G. Viral release was measured through the quantification of p24CA antigen concentration in the culture supernatants at 48 hours post-transfection. MonoMac6 cells were infected with VSV-G pseudotyped WT and ΔVpr viruses for 48 hours. (B) The VSV-G pseudotyped WT and ΔVpr stocks generated from pNL4-3Δenv-luc and pNL4-3ΔenvΔVpr-luc with/without aPKC inhibitor treatment were analyzed by immunoblotting for the incorporation of Vpr into virion particles. (C) Viral infectivity was detected by measuring the luciferase activity in the cell lysates. Data are mean ± s.e.m. of three independent experiments. (D) Effects of aPKC inhibitors on viability of MonoMac6 cells. MonoMac6 cells were treated with aPKC inhibitor (2 or 5 μM) or 1 μM Staurosporine (STS) and analyzed for cell viability using trypan blue exclusion at 72 hours. Data are mean ± s.e.m. of three independent experiments: **p < 0.01, Student’s t-test.