IRF-1 is induced by Nef.
(A) Western blot analysis of IRF-1 and β- tubulin levels on total cell extracts of MDMs 8 hours after the infection with either Δenv or Δenv/nef (VSV-G) pseudotyped HIV-1. (B) Western blot analysis of IRF-1 and β-tubulin levels on total MDM extracts after treatment with 100 ng/mL rNef for the indicated time intervals. Specific signals are indicated on the left side; molecular size markers (in kilodaltons) are reported on the right. (C) EMSA performed by using total cell extracts from either MDMs treated with 100 ng/mL rNef or HeLa cells treated with 500 IU/mL rIFN-α2b and the32P-labeled ISRE element of the human 2′-5′A synthetase promoter as a probe. Supershifts were performed with anti-STAT2, –IRF-1 or -p48 antibodies. Electrophoretic mobility of both ISGF-3/ISRE and IRF-1/ISRE complexes are indicated on the left side.
