LMP1 represses the core promoter activity of CD99 in a dose-dependent manner.

Relative luciferase activity was measured in 293 cells cotransfected with 0.5 μg CD99 promoter-driven luciferase construct p(–137/+123)luc, together with various amounts of the LMP1 expression plasmid (pcDNA3-LMP1). Transfected cells were prepared as previously described. After protein quantitation, lysates were separated by 10% SDS-PAGE and electroblotted onto the nitrocellulose filters. The same filter was probed with monoclonal antibodies such as CS1-4 and anti-Calnexin for LMP1 and Calnexin, respectively. One microgram pNF-κB CAT was cotransfected to confirm the activity of the cotransfected LMP1. Relative luciferase activity values represent mean ± SD of 3 independent transfections, whereas CAT values are the averages of 2 transfections among them.