FIG 3.

The IKK inhibitor BAY11 blocks vorinostat-mediated nuclear translocation of p65 and decreases NF-κB target gene expression. PC3 cells treated with VSV (MOI = 0.01) with or without vorinostat (5 μM) for 24 h were also pretreated with 10 μM BAY11 for 1 h where indicated. (A) Nuclear (N) and cytosolic (C) extracts were separated by gel electrophoresis and immunoblotted for RELA/p65. THOC1 was used as a loading control for the nuclear fraction. The numbers below the Western blots indicate nuclear RELA/p65/THOC1 signal ratios relative to the vehicle-treated cells ± standard errors. (B) Confocal microscopy analysis of PC3 cells that were infected with VSV-GFP (green), treated as indicated, and immunostained for RELA/p65 (red). (C) Relative mRNA levels of several NF-κB target genes in PC3 cells exposed to the indicated treatments and assessed by qPCR. White bars, vehicle (DMSO) treatment; light-gray bars, vorinostat treatment; dark-gray bars, VSV treatment; black bars, VSV-plus-vorinostat treatment; light-blue bars, VSV-plus-vorinostat-plus DMSO treatment; dark-blue bars, vorinostat-plus-VSV-plus BAY11 treatment. **, P ≤ 0.01; ***, P ≤ 0.005. The error bars indicate SD.