Figure 2

All Rel subunits are involved in TRAIL-induced NF-κB and apoptosis. (a) Whole-cell extracts of Panc1 cells transfected with the indicated siRNA for 48 h were submitted to western blotting for the indicated Rel subunit and Hsp90 as the loading control. A representative of four independent experiments with two different sets of siRNA is shown. (b) Cells were transfected with the indicated siRNA (set1) for 48 h, treated with 10 ng/ml TRAIL for 3 h and nuclear extracts were submitted to EMSA for measurement of NF-κB-binding activity. A representative of four independent experiments is shown. (c and d) Cells were transfected with with two different sets of siRNA (as indicated) for 48 h and then treated with 10 ng/ml TRAIL for 24 and 5 h, respectively. Apoptosis was determined by analysing sub-G1 content (c) or caspase-3/-7 activity (d). Data are presented as % of sub-G1 content over basal (untreated cells) or expressed as n-fold caspase-3/-7 activity normalized to the cellular protein content and represent the mean value±S.D. from six independent experiments. *P-values <0.05