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 2. The promoter of human PME-1 contains two putative LEF1/TCF elements. (A) Human PME-1 promoter region has two potential LEF1/TCF like elements (red). The promoter (-1000 to +389) of human PME-1 was analyzed by MatInspector software. Cis-elements are labeled with different color. There are two potential LEF1/TCF like elements. Other cis-elements are NFAT (nuclear factor of activated T cells), E2F (transcription factor family including E2F- and DP-like subunits), c-myb (Cellular homologue of avian myeloblastosis virus oncogene). (B) The SH-SY5Y cells were overexpressed with β-catenin tagged with HA. Monoclonal anti-HA were used to immunoprecipitate β-catenin and co-immunoprecipitated LEF1/TCF cis-elements were amplified by PCR using primers specific to LEF1/TCF cis-elements 1 and 2. (C) Schematic diagram of PME-1 promoter and its luciferase reporter plasmid. Human PME-1 promoter was inserted into pGL4.10 containing luciferase reporter gene to generate pGL4/PME-1-1000. (D) pGL4/PME-1-1000 and pGL4.10 showed in panel C were transfected into HEK-293T or SH-SY5Y cells. The Photinus pyralis luciferase activity and Renilla reniformis luciferase activity were measured subsequently and the Photinus pyralis luciferase activity was normalized with Renilla reniformis luciferase activity. Data are presented as mean ± SD (n=3); *P < 0.05, **P < 0.01.