Luciferase Assay for p53 Activity

A549 cells were cotransfected with p53-Luc plasmid (Agilent Technologies, Santa Clara, CA, USA) and pRL-TK Renilla vector (Promega, Madison, WI, USA) using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA). After 24 h, the transfected cells were treated with passive lysis buffer at room temperature for 10 min. Cell lysates were then transferred to a plate, and luciferase assay reagent and stop reagent were added in sequence. Next, luciferase activity was measured using a Luciferase Assay System (Promega, Madison, WI, USA) according to the manufacturer’s instructions, as previously described^{31 (link)}.

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Jiang W., Jin G., Cai F., Chen X., Cao N., Zhang X., Liu J., Chen F., Wang F., Dong W., Zhuang H, & Hua Z.C. (2019). Extracellular signal-regulated kinase 5 increases radioresistance of lung cancer cells by enhancing the DNA damage response. Experimental & Molecular Medicine, 51(2), 1-20.

Publication 2019

p53-Luc plasmid pRL-TK Renilla vector Lipofectamine 2000 reagent Luciferase Assay System

A549 cells Assay Buffer Cell Lipofectamine 2000 Luciferase Plasmid Promega Renilla Vector

Corresponding Organization : Changzhou University

Other organizations : Nanjing Medical University

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Variable analysis

independent variables

Treatment with passive lysis buffer

dependent variables

Luciferase activity

control variables

Transfection of p53-Luc plasmid and pRL-TK Renilla vector
Use of Lipofectamine 2000 reagent
Incubation time of 24 hours

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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