For regulation of IGF-2 constructs: shTbx15 and control C2C12 cells or 293FT cells at 70% confluence in D-MEM with 10% FBS grown to were co-transfected with 1 μg of a previously described of IGF-2 promoter constructs 2 (link), pBABE-Tbx15 or BABE-Empty, and pRenilla-TK (Promega), using Superfect transfection reagent (Quiagen). All transfections were harvested 24 h later and luciferase activity was measured using a Dual Renilla Luciferase II Assay Kit, and normalized to Renilla luciferase measurements (Promega).
Dual renilla luciferase 2 assay kit
Manufactured by Promega
The Dual Renilla Luciferase II Assay Kit is a laboratory tool designed to measure the activity of the Renilla luciferase reporter gene. It provides a sensitive and quantitative method for detecting and analyzing gene expression in various experimental systems.
Automatically generated - may contain errors
4 protocols using dual renilla luciferase 2 assay kit
1
Regulation of IGF-2 Expression via Tbx15
2
STAT5-Mediated Regulation of FSP27 Expression
3
Regulation of IGF-2 Expression by Tbx15
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For regulation of IGF-2 constructs: shTbx15 and control C2C12 cells or 293FT cells at 70% confluence in DMEM with 10% fetal bovine serum grown to were co-transfected with 1 μg of a previously described of IGF-2 promoter constructs30 (link), pBABE-Tbx15 or BABE-Empty and pRenilla-TK (Promega), using Superfect transfection reagent (Quiagen). All transfections were harvested 24 h later and luciferase activity was measured using a Dual Renilla Luciferase II Assay Kit, and normalized to Renilla luciferase measurements (Promega).
4
Regulation of G0S2 Luciferase Activity
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293T cells were transfected with polyethylenimine (PEI) transfection reagent while still in suspension in 96-well plates. Each well was transfected with 50 ng of a previously described G0S2 luciferase construct [33] (link), together with a total of 50 ng of expression plasmids containing STAT5A, STAT5B or empty vector controls and 10 ng of Renilla-TK plasmid. The cells were harvested 24 h later and luciferase activity was measured using a Dual Renilla Luciferase II Assay Kit and normalized to Renilla luciferase measurements (Promega). Site-directed mutagenesis of the G0S2 constructs by PCR was performed. The reporter vector was co-transfected with either a vector control or 25 ng of PPARγ expression vector and 25 ng of the obligate heterodimer RXRα.
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