An Epigenetic Compound Library Screen Identifies BET Inhibitors That Promote HSV-1 and -2 Replication by Bridging P-TEFb to Viral Gene Promoters through BRD4
Fig 5
HSV-1 infection induces protein complex formation and the association with viral gene promoters.
HeLa cells were infected with HSV-1 at 10 MOI for times as indicated. The samples were then processed for detection of protein association by immunoprecipitation assay, for protein co-localization by immunostaining, for protein-DNA interactions by a modified ChIP assay, respectively. (A) HSV-1 infection promotes protein association involving BRD4, CDK9, and RNAP II. Proteins in the lysates were used as control for input. The bar graph represents quantitative measurement of protein band intensities in anti-BRD4 immunocomplexes. The data are presented as mean ± SEM of 2 independent measurement. (B and C) Co-localization of BRD4 with CDK9 (B) and Rpb-1/RNAP II (C). The nuclei were stained using DAPI (blue). (D to G) Viral gene promoters in the BRD4 or Rbp-1/RNAP II immunocomplexes were determined by PCR (D, F) and real-time PCR (E, G). The GAPDH and IFNβ gene promoters in the complexes were included as controls for validation of ChIP assays.