Monitoring YAP Transcriptional Activity

YAP transcriptional activity was monitored via a luciferase reporter assay approach [35 (link)]. Briefly, Lipofectamine 3000 was used to transfect MDA-MB-231 cells with the 8xGTIIC-luciferase (Plasmid #34615) and β-gal plasmids (Ambion, USA). A Luciferase Reporter assay kit (BioVision, Inc., K801-200) was then used to monitor luciferase activity within these cells, while a β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer (Promega Corporation, E2000) was used based upon provided instructions to assess β-gal activity.

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She Q., Chen Y., Liu H., Tan J, & Li Y. (2022). A high level of the long non-coding RNA MCF2L-AS1 is associated with poor prognosis in breast cancer and MCF2L-AS1 activates YAP transcriptional activity to enhance breast cancer proliferation and metastasis. Bioengineered, 13(5), 13437-13451.

Publication 2022

Lipofectamine 3000 Luciferase Reporter assay kit β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer

Assay Buffer Cells Enzyme assay Lipofectamine Luciferase Mda mb 231 cells Plasmids Promega Transcriptional β galactosidase

Corresponding Organization : Baoji City Central Hospital

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Variable analysis

independent variables

Transfection of MDA-MB-231 cells with 8xGTIIC-luciferase (Plasmid #34615) and β-gal plasmids

dependent variables

Luciferase activity
β-Galactosidase (β-gal) activity

control variables

Lipofectamine 3000 used for transfection
Luciferase Reporter assay kit (BioVision, Inc., K801-200) used to monitor luciferase activity
β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer (Promega Corporation, E2000) used to assess β-gal activity

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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