Overexpression of natural antisense BCMA decreases BCMA-mediated NF-kappaB activation. 293 cells were co-transfected with a total of 1 μg of various plasmids. All samples contained 100 ng of the luciferase reporter plasmid and 50 ng of pGKβ-gal plasmid. The negative control value was obtained the insert-less pcDNA3 vector (vector). NF-kappaB activation was measured in samples transfected with a) 100 ng of pcDNA3aSp12 (p12), b) 100 ng of pcDNA3LMP1 (LMP1), c) 100 ng each of LMP1 and p12 (LMP1+p12), d) 100 ng of pcDNA3hBCMA (hBCMA) and e) 100 ng each of hBCMA and p12 (hBCMA+p12). The quantity of plasmid added was adjusted to 1 μg by adding sufficient insert-less pcDNA3 vector. 18–24 h after transfection, cells were harvested, lysed and analysed for luciferase and β-galactosidase activity. Luciferase values were normalised to β-galactosidase activity and the results are displayed as a multiple of the induction by vector alone. A representative result of three independent experiments is shown. Error bars denote standard deviations for triplicate samples.