Hypoxia and VEGF Promoter Luciferase Assay

HIF-1α binding hypoxia-response element (HRE) and VEGF promoter containing luciferase vector (HRE-Luc and VEGF-Luc) were kind gifts from Dr. Jong-Wan Park (Seoul National University) [38 (link)]. For luciferase assay, luciferase and β-gal vector were transiently transfected into HEK293 or Caki-1 cells using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) and PolyFect transfection reagent (Qiagen, Venlo, Netherlands). After hypoxia or drug treatment in transfected cells, luciferase activities were measured using a Synergy 2 Luminometer (BioTek, Winooski, VT, USA) and normalized by β-gal assay.

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Lee Y.M., Kim G.H., Park E.J., Oh T.I., Lee S., Kan S.Y., Kang H., Kim B.M., Kim J.H, & Lim J.H. (2019). Thymoquinone Selectively Kills Hypoxic Renal Cancer Cells by Suppressing HIF-1α-Mediated Glycolysis. International Journal of Molecular Sciences, 20(5), 1092.

Publication 2019

Lipofectamine 2000 PolyFect transfection reagent Synergy 2 Luminometer

Assay Cells Drug Gifts Hypoxia Lipofectamine 2000 Luciferase Response element Transfection Vector Vegf

Corresponding Organization : Konkuk University

Other organizations : Yonsei University, Korea University

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Variable analysis

independent variables

Hypoxia treatment
Drug treatment

dependent variables

Luciferase activity
β-gal activity

control variables

Transfection of luciferase and β-gal vector into HEK293 or Caki-1 cells
Use of Lipofectamine 2000 and PolyFect transfection reagents

controls

Positive control: HRE-Luc and VEGF-Luc vectors
Negative control: Not explicitly mentioned

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