Research Paper Volume 11, Issue 21 pp 9672—9688

Glycogen synthase kinase-3β suppresses the expression of protein phosphatase methylesterase-1 through β-catenin

Figure 5. PME-1 expression is regulated by GSK-3β/β-catenin pathway. (A, B) HEK-293T cells were transiently transfected with GSK-3β. Levels of PME-1, β-catenin, GSK-3β, and Non-pS-β-catenin (dephosphorylated β-catenin at Ser33, Ser37 and Thr41) were analyzed by Western blots and normalized with GAPDH or corresponding proteins (B). (C, D) HA-tagged GSK-3β was overexpressed in HeLa cells and immuno-stained by polyclonal rabbit anti-β-catenin or mouse monoclonal anti-HA (GSK-3β) followed by florescence labeled anti-rabbit (green) or anti-mouse (red) second antibodies, respectively (C). The fluorescence levels of the nucleus and the cytoplasm were measured by IMAGE J and nucleus/cytoplasm ratio of β-catenin was analyzed (D). (E) pGL4/PME-1-1000 were co-transfected with GSK-3β or siGSK-3β in HEK-293T cells. The luciferase activity was measured. (F) HEK-293T cells were co-transfected with β-catenin and/or GSK-3β with pGL4/PME-1-1000. The luciferase activity was measured. *: compared with control (Con). &: compared with β-catenin. (G) The luciferase activity as measured. *: compared with control (Con), #: compared with β-catenin. Data are presented as mean ± SD (n=3), *P < 0.05, **P < 0.01, ***P < 0.001.