Luciferase Assay for Transcriptional Activity

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 (47 (link)). Cells were transfected with 100 ng HOP-Flash (Catalog # 83467, Addgene) or HIP-Flash luciferase reporter plasmid (Catalog # 83466, Addgene), plus 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., Fu Q., Bian X., Fu Y., Xin J., Liang N., Li S., Zhao Y., Fang L., Li C., Zhang J., Dionigi G, & Sun H. (2021). Long Non-Coding RNA MAPK8IP1P2 Inhibits Lymphatic Metastasis of Thyroid Cancer by Activating Hippo Signaling via Sponging miR-146b-3p. Frontiers in Oncology, 10, 600927.

Publication 2020

HOP-Flash HIP-Flash luciferase reporter plasmid pRL-TK Renilla plasmid Lipofectamine 3000 Dual Luciferase Reporter Assay Kit

Assay Cells Lipofectamine Luciferase Plasmid Promega Renilla Transfection

Corresponding Organization :

Other organizations : Union Hospital, Jilin University, University of Messina

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Variable analysis

independent variables

Transfection with HOP-Flash luciferase reporter plasmid
Transfection with HIP-Flash luciferase reporter plasmid

dependent variables

Luciferase signal
Renilla signal

control variables

Cell number (4 × 10^4 cells)
Culture duration (24 h)
Transfection with pRL-TK Renilla plasmid

controls

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

Annotations

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