Figure 1
GSK-3β is involved in NF-κB activation.
(a) HEK293 cells were transfected either with control siRNA (siCon) or with siRNA-targeting GSK-3β (siGSK-3β) or siRNA-targeting GSK-3α (siGSK-3α). After 24 h stimulation with TNFα (1ng/ml), NF-κB-dependent gene expression was quantified by measurement of the luciferase activity. Fold induction is the ratio of stimulated to unstimulated cells (**P < 0.01). Cell lysates were immunoblotted with the antibodies indicated. (b) HEK293 cells were stably transfected either with control shRNA (shCtr) or with shRNA-targeting GSK-3β (shGSK-3β). After 24 h stimulation with TNFα (1ng/ml), NF-κB-dependent gene expression was quantified by measurement of the luciferase activity. Cell lysates were immunoblotted with the antibodies indicated. (c) In vitro kinase assay using recombinant human 1 μg NEMO as substrate for GSK-3β (0.01 μg or 0.05 μg). (d) GSK-3β (0.01 μg) was incubated with 1 μg recombinant human NEMO and 10 μCi of [γ-32P]ATP at 30 °C for different time periods.
