Luciferase reporter assay in cells

The K1 and B-CPAP cells (4 × 10⁴) were seeded in triplicate in 24-well plates and cultured for 24 h, and the luciferase reporter assay was performed as previously described (Ren et al. 2019 (link)). Cells were transfected with 250 ng (CAGAC) 12/pGL3 reporter luciferase plasmid or 5 ng pRL-TK Renilla plasmid (Promega) using Lipofectamine 3000 (Invitrogen) according to the manufacturer’s recommendation. Luciferase and Renilla signals were measured 36 h after transfection using a Dual Luciferase Reporter Assay Kit (Promega) according to the manufacturer’s protocol.

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Liu X., Zhang C., Wang X., Cui C., Cui H., Zhu B., Chen A., Zhang L., Xin J., Fu Q., Dionigi G, & Sun H. (2023). Long non-coding RNA MFSD4A-AS1 promotes lymphangiogenesis and lymphatic metastasis of papillary thyroid cancer. Endocrine-Related Cancer, 30(3), e220221.

Publication 2023

pRL-TK Renilla plasmid Lipofectamine 3000 Dual Luciferase Reporter Assay Kit

Assay B cells Cells Cpap Lipofectamine Luciferase Pgl3 Plasmid Promega Renilla Transfection

Corresponding Organization :

Other organizations : Union Hospital, Jilin University, University of Milan, IRCCS Istituto Auxologico Italiano

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Variable analysis

independent variables

Cell type (K1 and B-CPAP cells)

dependent variables

Luciferase reporter activity

control variables

Cell seeding density (4 × 10^4 cells)
Culture duration (24 hours)
Transfection conditions (250 ng (CAGAC)12/pGL3 reporter luciferase plasmid and 5 ng pRL-TK Renilla plasmid using Lipofectamine 3000)
Measurement timing (36 hours after transfection)
Measurement method (Dual Luciferase Reporter Assay Kit)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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