(A–J) FANCD2 (A–E) and FANCI (F–J) are required for HIV-1 DNA integration. Indicated cells were infected with VSV-G pseudotyped HIV-Luc virus. The expression of indicated proteins was determined by western blot (A and F). Luciferase activities (B and C), total viral DNA (C and H), unintegrated viral DNA (D and I), and integrated viral DNA (E and J) were measured at indicated time points post-infection. Data are represented as means ± SDs from 3 independent experiments (n = 3). ns p > 0.05; *p < 0.05; **p < 0.01.
(K and L) Knockdown of FANCI inhibits the replication of HIV-1. MT-4 cells stably expressing control shRNA (SCR) or shRNAs targeting FANCI were infected with HIV-1(NL4.3) at 1 ng CA/106 cells. The expression of FANCI was determined by western blot (K). Viral spreading was monitored by assay for the infectivity of supernatants in TZM-bl cells engineered with the HIV-1 Tat-responsive luciferase reporter gene (L). Data presented are means ± SEMs from 2 technical replicates and are representative of 2 independent experiments.
See also Figure S2.