Figure 5.

GSK-3β differentially affects the binding of NF-κB p65 and p50 to the BCL2L1 and BIRC3 promoters. (a, b) qRT-PCR analysis of MEF and Panc04.03 cells following TNFα-stimulation. Note that Bcl-xL and cIAP2 transcripts, but not IKBα, were reduced in Gsk-3β-null MEFs compared with WT control (a). Similarly, Bcl-xL and cIAP2, but not IKBα mRNA expression was reduced in TNFα-treated Panc04.03 cells transduced with lentiviral GSK-3β shRNA (b). (c–e). ChIP analysis of Panc04.03 cells treated with GSK-3i (0.5 μM LY2064827) or diluent (0.1% DMSO) for 16 h. The relative amount of DNA precipitated with the specified antibodies was accessed by qPCR using three sets of primers targeting promoter regions containing the depicted NF-κB sites. The qPCR was performed in triplicate; and the result is normalized to diluent control