Stable cell lines expressing the TOP-FLASH reporter were generated by transducing
cells with 7TFP recombinant lentiviruses (Fuerer
and Nusse, 2010 ), and luciferase assay was performed as previously
described (Singh et al., 2012 (link)). Briefly,
Rediject D-Luciferin Ultra (Perkin Elmer) was added in 0.2 ml fresh media
(1–200 dilution) to each well of cells in a 96-well plate and incubated for 15
min at 37°C. Luciferase activity was imaged with the IVIS Lumina II In Vivo
Imaging System (Perkin Elmer). The radiance of each well was determined using Living
Image 4.2 software (Perkin Elmer), background corrected by subtracting the mean
signal from empty wells and normalized both to the relative cell number of each well
as determined by Syto60 assay and the resulting normalized mean value of untreated
wells.
cells with 7TFP recombinant lentiviruses (
and Nusse, 2010
described (Singh et al., 2012 (link)). Briefly,
Rediject D-Luciferin Ultra (Perkin Elmer) was added in 0.2 ml fresh media
(1–200 dilution) to each well of cells in a 96-well plate and incubated for 15
min at 37°C. Luciferase activity was imaged with the IVIS Lumina II In Vivo
Imaging System (Perkin Elmer). The radiance of each well was determined using Living
Image 4.2 software (Perkin Elmer), background corrected by subtracting the mean
signal from empty wells and normalized both to the relative cell number of each well
as determined by Syto60 assay and the resulting normalized mean value of untreated
wells.
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