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. Author manuscript; available in PMC: 2010 Aug 12.

Published in final edited form as: Oncogene. 2009 Sep 28;29(1):128–138. doi: 10.1038/onc.2009.302

Over-expression of Pdcd4 up-regulates E-cadherin and inhibits β-catenin/Tcf dependent transcription. (a) Over-expression of Pdcd4 in LoVo cells up-regulates E-cadherin expression. Western blot was performed using antibodies against E-cadherin, Pdcd4, and GAPDH. The ratios of E-cadherin/GAPDH and Pdcd4/GAPDH in LoVo-control cells are designated as 1. The number indicates the relative level of either E-cadherin or Pdcd4 protein. (b) Pdcd4 inhibits β-catenin/Tcf dependent transcription. TOPflash (0.2 μg, filled bar) and FOPflash (0.2 μg, open bar) along with 10 ng of pRL-SV40 were transfected into LoVo-control and LoVo-Pdcd4 cells. The luciferase activity was normalized against Renilla activity. The activity of LoVo-control cells transfected with TOPflash and FOPflash is designated as 100%. The asterisk denotes a significant difference compared with cells transfected with TOPflash as determined by one-way ANOVA (P < 0.005). (c) Pdcd4 inhibits β-catenin/Tcf dependent transcription in a dose-dependent manner. Increasing amount (0-1.5 μg) of pcDNA-Pdcd4 plasmid was transfected along with 0.2 μg of TOPflash (filled bar) or FOPflash (open bar) into LoVo cells. The luciferase activity was normalized against Renilla activity. The total DNA was maintained at 1.7 μg by adding empty vector pcDNA3.1 DNA. The activity of cells transfected with TOPflash or FOPflash plasmid but without pcDNA-Pdcd4 plasmid is designated as 100%. The asterisk denotes a significant difference compared with cells transfected with TOPflash but without pcDNA-Pdcd4 plasmid, as determined by one-way ANOVA (P < 0.005). The experiments in (b) and (c) were repeated three times each with five independent transfections and representative data are shown. Results are expressed as mean ± SD.

Over-expression of Pdcd4 up-regulates E-cadherin and inhibits β-catenin/Tcf dependent transcription. (a) Over-expression of Pdcd4 in LoVo cells up-regulates E-cadherin expression. Western blot was performed using antibodies against E-cadherin, Pdcd4, and GAPDH. The ratios of E-cadherin/GAPDH and Pdcd4/GAPDH in LoVo-control cells are designated as 1. The number indicates the relative level of either E-cadherin or Pdcd4 protein. (b) Pdcd4 inhibits β-catenin/Tcf dependent transcription. TOPflash (0.2 μg, filled bar) and FOPflash (0.2 μg, open bar) along with 10 ng of pRL-SV40 were transfected into LoVo-control and LoVo-Pdcd4 cells. The luciferase activity was normalized against Renilla activity. The activity of LoVo-control cells transfected with TOPflash and FOPflash is designated as 100%. The asterisk denotes a significant difference compared with cells transfected with TOPflash as determined by one-way ANOVA (P < 0.005). (c) Pdcd4 inhibits β-catenin/Tcf dependent transcription in a dose-dependent manner. Increasing amount (0-1.5 μg) of pcDNA-Pdcd4 plasmid was transfected along with 0.2 μg of TOPflash (filled bar) or FOPflash (open bar) into LoVo cells. The luciferase activity was normalized against Renilla activity. The total DNA was maintained at 1.7 μg by adding empty vector pcDNA3.1 DNA. The activity of cells transfected with TOPflash or FOPflash plasmid but without pcDNA-Pdcd4 plasmid is designated as 100%. The asterisk denotes a significant difference compared with cells transfected with TOPflash but without pcDNA-Pdcd4 plasmid, as determined by one-way ANOVA (P < 0.005). The experiments in (b) and (c) were repeated three times each with five independent transfections and representative data are shown. Results are expressed as mean ± SD.