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RelA/p65 Ser-468 is a suitable direct substrate for GSK3β. A, sequence analysis indicates that Ser-468 but not Ser-276 is located in a GSK3β consensus motif. B, GSK3β directly and efficiently phosphorylates RelA/p65 at Ser-468. Glutathione S-transferase (GST)-conjugated recombinant GSK3β and p65 were prepared for in vitro kinase assay as indicated. Total cell lysates extracted from TNF-α-stimulated HKC cells served as a positive control for different molecules probed.
