Expression of antisense RNA is sufficient to obtain inhibition. A. 293T cells were co-transfected with a total of 1 μg of various plasmids, using the same conditions for measuring NF-kappaB activation as in Fig. 1. The negative control was insert-less SV40 promoter-driven pSG5FLAG vector (vector). All other samples were transfected with 100 ng of pSG5FLAGhBCMA (FhBCMA) vector and various amounts of pcDNA3aSp12 (p12) or pcDNA3aSp12-ATG (p12-ATG), lacking the initiation ATG codon. Cells were harvested and lysed 48 hours after transfection and the lysate was assayed for NF-kappaB activation. B. Lysate (20 μg) were subjected to electrophoresis and the amount of FhBCMA in 293T cells was determined by western blotting using the M2 anti-FLAG monoclonal antibody. We checked that the amount of protein was similar in all samples of 293T cells by subjecting 10 μg of cell lysate to western blotting using a rabbit polyclonal anti-PI3-Kp85 antibody. C. The positive control was transfection with either 100 ng of LMP1 or 100 ng of LMP1 and 750 ng of p12 or p12-ATG expression vector.