Lentivirus-expressed nef shRNA inhibits HIV-1 replication and affects chemokine production in MDMs. (A) MDMs were transduced with Lenti cont or Lenti shNef366 at an MOI of 2. At 2 hours after infection, cells were washed twice, then cultured for another 24 hours, at which point the cells were infected with HIV-1_NF462. The culture supernatants were collected at 3- or 4-day intervals after HIV infection, and the level of p24 antigen was measured by ELISA. (B) MDMs transduced either with Lenti control or Lenti shNef366 were infected with HIV-1 and supernatants were collected 10 days after infection and designated as HIV-1 Lenti cont and HIV-1 Lenti shNef366, respectively, CEMx174 CCR5/LTR-EGFP cells were infected either with HIV-1 Lenti cont or HIV-1 Lenti shNef366 and GFP⁺, HIV-1–infected T cells were analyzed by FACS 48 hours later. The data represent the average ± SD of 3 independent experiments. (C) The culture supernatants of MDMs transduced with lentivirus vectors were collected 14 days after infection and the levels of the chemokines MIP-1α and MIP-1β were measured.