SERCA2 regulates NF-κB promoter activity and IL-8 release.
NF-κB luciferase reporter-expressing 16HBEo- cells were cultured on collagen-coated 6-well plates and transfected with either control siRNA or SERCA2 siRNA. Forty-eight hour post transfection cells were treated with ‘Cytomix’ containing TNFα (20 ng/ml), IL-1β (10 ng/ml), IFNγ (10 ng/ml) and LPS (50 ng/ml) with or without thapsigargin (thaps; 2 µM)). (A) Cell lysates were prepared 24 h after treatment and luciferase activity was measured as described. The data shown are mean±SEM (n = 6). The image is representative of four independent experiments. * Indicates significant difference from untreated siControl transfected cells, # indicates significant difference from untreated siSERCA2 transfected cells, $ indicates significant difference from cytomix-treated siControl transfected cells, and $$ indicates significant difference from cytomix+thaps-treated siControl-transfected cells p<0.05. (B) Cells were cultured and treated as described above and supernatant media was collected and analyzed for IL-8. The data shown are mean±SEM (n = 3). The image is representative of two independent experiments. * Indicates significant difference from untreated siControl transfected cells, # indicates significant difference from untreated siSERCA2 transfected cells, $ indicates significant difference from cytomix-treated siControl transfected cells and $$ indicates significant difference from cytomix+thaps-treated siControl-transfected cells p<0.05.