FIGURE 3.

Forced expression of a mutant uninhibitable GSK3β (S9A) blocks the suppressive effect of HGF on TNF-α induced phosphorylation of p65 at Ser-468. A, HKC cells were transiently transfected with the pcDNA3 EV or the vector encoding the HA-conjugated wild type GSK3β or the mutant GSK3β in which the Ser-9 was replaced by alanine. Whole cell lysates were harvested and analyzed for different molecules by Western immunoblot. Fluorescent immunocytochemistry staining of HA demonstrated that ∼70% cells expressed the vector. B, after they were transfected with different vectors, HKC cells were subjected to different treatments as indicated. Whole cell lysates underwent immunoblot assay. C, densitometric analysis of the immunoblot in B shows that the inhibitory effect of HGF on TNF-α-induced phosphorylation of p65 at Ser-468 is obliterated in HKC cells expressing GSK3β S9A. *, p < 0.05 versus TNF-α-alone-treated cells with the same transfection.