Mouse L cells stably expressing SuperTOPflash reporter (Mikels and Nusse 2006 (link)) were cultured in DMEM, 10% fetal bovine serum (FBS), and antibiotics. L cells were plated into a 24-well plate, allowed to recover for 5–8 h, and then treated in duplicate or triplicate with Wnt3A, Wnt4, Wnt7B, Rspo1, or Dkk1 proteins, conditioned medium, or lentivirus as described in the text. Luciferase assays were performed using the Dual-Light reporter gene assay system (Applied Biosystems). Relative luciferase units were measured and normalized against β-galactosidase activity at 48 h post-treatment.
Protocol detail
Wnt Signaling Pathway Activation Assay
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Antibiotics
Assay
Conditioned medium
Fetal bovine serum
L cells
Lentivirus
Light
Luciferase
Mouse
Proteins
Reporter gene
Wnt4
β galactosidase
Corresponding Organization :
Other organizations : Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai Institutes for Biological Sciences
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Variable analysis
independent variables
- Wnt3A
- Wnt7B
- Rspo1
- Conditioned medium
- Lentivirus
- Luciferase activity
- β-galactosidase activity
- L cells stably expressing SuperTOPflash reporter
- DMEM culture medium
- 10% fetal bovine serum (FBS)
- Antibiotics
- Incubation time (48 h)
- Not specified
- Not specified
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