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Inhibition of Wnt signaling by spiperone . HEK293 cells were cotransfected with a TOPflash reporter construct, along with vectors for: (A) control (pcDNA3 plasmid alone), Wnt1/LRP6, or Wnt3/LRP6; (B) control (pcDNA3 plasmid alone) or Dvl; (C) control (pcDNA3 plasmid alone) or β-catenin. (D) HEK293 cells were transfected with an <t>NFAT-Luc</t> reporter and an expression plasmid for NFATc. (E) HEK293 cells were transfected with an <t>AP1-Luc</t> reporter and an expression plasmid for H-Ras V12 . After transfection for 24 h, the cells were treated with or without spiperone (5 μM) for another 24 h, and then harvested, and extracted for determination of luciferase activities. The β-galactosidase control plasmid was used to correct for transfection efficiency. The results are expressed as fold induction of luciferase activity normalized to a β-galactosidase control, and are the means of three experiments ± SEM.
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Inhibition of Wnt signaling by spiperone . HEK293 cells were cotransfected with a TOPflash reporter construct, along with vectors for: (A) control (pcDNA3 plasmid alone), Wnt1/LRP6, or Wnt3/LRP6; (B) control (pcDNA3 plasmid alone) or Dvl; (C) control (pcDNA3 plasmid alone) or β-catenin. (D) HEK293 cells were transfected with an <t>NFAT-Luc</t> reporter and an expression plasmid for NFATc. (E) HEK293 cells were transfected with an <t>AP1-Luc</t> reporter and an expression plasmid for H-Ras V12 . After transfection for 24 h, the cells were treated with or without spiperone (5 μM) for another 24 h, and then harvested, and extracted for determination of luciferase activities. The β-galactosidase control plasmid was used to correct for transfection efficiency. The results are expressed as fold induction of luciferase activity normalized to a β-galactosidase control, and are the means of three experiments ± SEM.
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Inhibition of Wnt signaling by spiperone . HEK293 cells were cotransfected with a TOPflash reporter construct, along with vectors for: (A) control (pcDNA3 plasmid alone), Wnt1/LRP6, or Wnt3/LRP6; (B) control (pcDNA3 plasmid alone) or Dvl; (C) control (pcDNA3 plasmid alone) or β-catenin. (D) HEK293 cells were transfected with an NFAT-Luc reporter and an expression plasmid for NFATc. (E) HEK293 cells were transfected with an AP1-Luc reporter and an expression plasmid for H-Ras V12 . After transfection for 24 h, the cells were treated with or without spiperone (5 μM) for another 24 h, and then harvested, and extracted for determination of luciferase activities. The β-galactosidase control plasmid was used to correct for transfection efficiency. The results are expressed as fold induction of luciferase activity normalized to a β-galactosidase control, and are the means of three experiments ± SEM.

Journal: BMC Pharmacology

Article Title: Spiperone enhances intracellular calcium level and inhibits the Wnt signaling pathway

doi: 10.1186/1471-2210-9-13

Figure Lengend Snippet: Inhibition of Wnt signaling by spiperone . HEK293 cells were cotransfected with a TOPflash reporter construct, along with vectors for: (A) control (pcDNA3 plasmid alone), Wnt1/LRP6, or Wnt3/LRP6; (B) control (pcDNA3 plasmid alone) or Dvl; (C) control (pcDNA3 plasmid alone) or β-catenin. (D) HEK293 cells were transfected with an NFAT-Luc reporter and an expression plasmid for NFATc. (E) HEK293 cells were transfected with an AP1-Luc reporter and an expression plasmid for H-Ras V12 . After transfection for 24 h, the cells were treated with or without spiperone (5 μM) for another 24 h, and then harvested, and extracted for determination of luciferase activities. The β-galactosidase control plasmid was used to correct for transfection efficiency. The results are expressed as fold induction of luciferase activity normalized to a β-galactosidase control, and are the means of three experiments ± SEM.

Article Snippet: The NFAT-Luc and AP1-Luc reporters were purchased from BD Biosciences.

Techniques: Inhibition, Construct, Plasmid Preparation, Transfection, Expressing, Luciferase, Activity Assay