Wnt Signaling Reporter Assays

Reporter assays have been described earlier [30] (link). While LS/L cells [24] (link) had been stably transfected with pSuperTOPFlash and pEF1/Myc-His/LacZ (Invitrogen), HEK293A and HEK293T cells were transfected with pMegaTOPFlash [28] (link) and pEF1/Myc-His/LacZ or pEF1α-LacZ-IRES-Puro^R one day prior to the addition of Wnt3A. Reporter assays were performed with the dual-light combined reporter gene assay system (Applied Biosystems) with a Centro LB960 luminometer (Berthold) according to the manufacturer's instructions. β-galactosidase activity was used to normalize the values, which are represented with their standard deviation.

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Fuerer C, & Nusse R. (2010). Lentiviral Vectors to Probe and Manipulate the Wnt Signaling Pathway. PLoS ONE, 5(2), e9370.

Publication 2010

Assay Cells Ires L cells Lacz Light Reporter gene β galactosidase

Corresponding Organization :

Other organizations : Stanford University

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Variable analysis

independent variables

The type of cell line used: LS/L cells, HEK293A cells, and HEK293T cells

dependent variables

Luciferase activity from the reporter constructs (pSuperTOPFlash, pMegaTOPFlash)

control variables

The reporter construct pEF1/Myc-His/LacZ or pEF1α-LacZ-IRES-PuroR, which was used to measure β-galactosidase activity for normalization

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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