Figure 1

PKR is activated after HIV-1 infection and inhibited during active HIV-1 replication. A) HIV-1 pNL4-3 kinetics from infected PBMCs. 6.5 × 10⁷ PBMCs from a healthy donor were infected with HIV-1 pNL4-3. At day 7, cells were separated in two flasks and IFN α/β (10000U/mL) was added to the cells in one of them up to day 14. Aliquots of cell supernatant were collected at different times and assayed for RT activity. B) Protein expression of pNL4-3-infected PBMCs. 50 μg of whole-cell extracts from pNL4-3-infected PBMCs from different harvest times were subjected to a 10% SDS PAGE and blotted with anti-P-PKR, anti-PKR, anti-HIV-p24, anti-PACT, anti-ADAR1 and anti-actin antibodies as indicated. C) Protein expression of mock-infected PBMCs. 6.5 × 10⁷ PBMCs from the same donor as in B were cultured and passed at the same time as in B. IFN α/β was added similarly from day 7 to 14. 50 μg of whole-cell extracts from mock-infected PBMCs from the indicated times were subjected to a 10% SDS PAGE and blotted with anti-P-PKR, anti-PKR and anti-actin antibodies as indicated.