FIG. 7.

IE86 antibody recognizes the IR1-DNA complex-bound IE86 protein. (A) U373-IE86 nuclear extract was incubated with labeled IR1 oligonucleotide and either a monoclonal antibody specific for the HCMV IE proteins (MAb 810) or polyclonal antibodies specific for NF-κB p65 and p50 subunits (p65 Ab and p50 Ab, respectively). U373-IE86 with the IR1 probe was used as a control. (B to D) EMSA was done as a control (see panel A, lane 1), and then the gel was blotted onto DEAE (B) and nitrocellulose (C and D) membranes. The nitrocellulose membranes were subsequently probed with a monoclonal antibody specific for IE86 (see Materials and Methods). U373-IE86 extracts and recombinant IE86 protein (rIE86) alone were used as controls in EMSA-Western blot analysis (D). Arrows indicate the IR1 DNA-binding complex (A and B) and IE86 protein present in the IR1 DNA-binding complex (C and D).