The target gene of miR-128/miR-142 was predicted by TargetScan online prediction software. Based on the bioinformatics prediction results, partial HOXA10 3ʹUTR sequence containing wild-type or mutant-type miR-128/miR-142 targeting sites were amplified and cloned into pmirGLO vector (Promega, Madison, Wisconsin, USA), termed WT-HOXA10-3ʹUTR and MUT-HOXA10 reporter plasmids. Then, these constructed plasmids were cotransfected with miR-control, miR-128 or miR-142 into SCC-9 or SSC-25 cells and incubated for 48 h. Finally, dual-luciferase reporter assays kit (Promega) was used to detect the luciferase activity in cell lysates.
Dual luciferase reporter assays kit
Manufactured by Promega
Sourced in United States
The Dual-Luciferase Reporter Assay System is a laboratory tool that allows for the quantitative measurement of two different luciferase reporter enzymes within a single sample. It provides a method for analyzing gene expression or the activity of regulatory sequences.
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Lab products found in correlation
4 protocols using dual luciferase reporter assays kit
1
Validating miR-128/miR-142 Regulation of HOXA10
2
Dual Luciferase Assay for Wnt Signaling
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A pGL3 reporter containing target regions was transfected into breast cancer cells using a Renilla luciferase vector. Firefly and Renilla luciferase activities were detected using a Dual Luciferase Reporter Assays Kit (Promega, E1910, WI, USA) at 48 h post-transfection according to the manufacturer’s instruction.
TOPFlash/FOPFlash assay was performed in the same way. TOPFlash contains three copies of the TCF-4 binding sites, and FOPFlash (the control reporter) contains mutated TCF-4 binding sites. Renilla luciferase expression plasmid acted as the system control.
TOPFlash/FOPFlash assay was performed in the same way. TOPFlash contains three copies of the TCF-4 binding sites, and FOPFlash (the control reporter) contains mutated TCF-4 binding sites. Renilla luciferase expression plasmid acted as the system control.
3
Dual-Luciferase Assay for Breast Cancer
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A pGL3 reporter containing target regions was transfected into breast cancer cells using a Renilla luciferase vector. Firefly and Renilla luciferase activities were detected using a Dual-Luciferase Reporter Assays Kit (#E1910, Promega, Madison, WI, USA) at 48 h post-transfection according to the manufacturer’s instructions.
The TOP/FOP Flash assay was performed in the same manner. TOP Flash contains three copies of the TCF-4 binding sites, and FOP Flash (the control reporter) contains mutated TCF-4 binding sites. The Renilla luciferase expression plasmid acted as the system control.
The TOP/FOP Flash assay was performed in the same manner. TOP Flash contains three copies of the TCF-4 binding sites, and FOP Flash (the control reporter) contains mutated TCF-4 binding sites. The Renilla luciferase expression plasmid acted as the system control.
4
Dual-Luciferase Assay for ERRγ and PPARδ
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ERRγ and PPARδ overexpressing cells were produced as previously described and treated with VF [24 (link)]. After 24 h incubation, the luciferase activity was measured using the Dual-luciferase reporter assays kit (Promega, Madison, Wisconsin, USA) with Tecan microplate reader (Tecan) following the manufacturer's description.
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