Figure 7
From: The PKR activator, PACT, becomes a PKR inhibitor during HIV-1 replication
PACT interacts with ADAR1. A) Transfected Flag-PACT interacts with endogenous ADAR1. HEK 293T cells were not transfected (lanes 1, 3, 5), or transfected with 2 μg of pCMV2-Flag-PACT (lanes 2, 4, 6). IP was performed with 2 mg of protein and anti-Flag antibody and treated with Benzonase (lane 5, 6). 150 μg of cell lysate (input or immunoprecipitates) were resolved by 10% SDS-PAGE and analyzed by immunoblotting using antibodies against ADAR1, PACT and actin. B) Transfected ADAR1-V5 interacts with endogenous PACT. HEK 293T cells were not transfected (lanes 1, 3, 5), or transfected with 2 μg of pcDNA3.1-ADARp150-V5 (lanes 2, 4, 6). IP was performed with 2 mg of protein and anti-V5 antibody and treated with Benzonase (lane 5, 6). 150 μg of cell lysate (input or immunoprecipitates) were resolved by 10% SDS-PAGE and analyzed by immunoblotting using antibodies against ADAR1, PACT and actin.