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Examples
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Plasmids for mouse GST-ARE-dependent firefly luciferase, control renilla luciferase (hRluc / TK from Promega), and other genes were co-transfected into MDA-MB-231 cells, followed by treatment with indicated chemicals for 16 hr.
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MDA-MB-231 cells transfected with plasmids for ARE-driven firefly luciferase, TK-renilla luciferase and the indicated genes were treated with 7.5 µM SF or 50 µM tBHQ for 16 hr prior to cell lysis.
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Firefly luciferase activity was normalized according to the activity of renilla luciferase.
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Skov-3 tumor cells were stably transduced using a lentivirus expressing red fluorescent protein and renilla luciferase (rLuc)
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Values were normalized for transfection efficiency according to the activity of a co-transfected renilla luciferase plasmid and expressed relative to the activity of pGL3-basic vector.
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For every sample, both the ratio between the luminescence of firefly and renilla luciferases and the firefly luciferase activity per µg of protein in cell lysates were obtained.
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The ratio between the activity of firefly and renilla luciferases is plotted.
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To test whether the phosphorylation of CREB described above could lead to gene transcription through its cognate activator sequence cAMP response element (CRE), Jurkat cells were double-transfected with two plasmids, one coding for firefly luciferase under the control of CRE and the other coding for renilla luciferase under the control of a viral promoter with constitutive transcriptional activity.
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Stimulation of pCREB transcriptional activity in the transfected cells will result in an increase of the ratio of firefly luciferase activity to that of renilla luciferase activity.
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The ratio between the activity of firefly and renilla luciferases is plotted.
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