Overexpression of BRD4 mutants on HSV-1 infection.

(A) Schematic drawing of BRD4 constructs. The DNA was subcloned into p3xFLAG-CMV-24 vector for mammalian cell expression. (B) BRD4 wt and its mutants were detected in the nuclei of transfected 293T cells. (C) Transient transfection of BRD4 wt had no effect on HSV-1 infection, determined by ICP0 and ICP4 expression. 293T cells in 6-well plates were transfected with p3xFLAG empty vector or with p3xFLAG-BRD4 wt at 0.3 and 1.0 μg for 24 hr. The cells were then infected with HSV-1 at 0.5 MOI for 24 hr. (D) Transient transfection of BD1, BD1/2 or ΔBD1 increases HSV-1 ICP0 and ICP4 expression. 293T cells were transfected and used as in (C). (E) BRD4 mutants promote HSV-1 and HSV-2 infection as were determined by plaque forming assay.