Transcriptional Activity Measurement

AR transcriptional activity was determined by ARE-luciferase (ARR3tk-eGFP/SV40-mCherry; 132360; Addgene, Watertown, MA, USA) [23 (link)]. ER transcriptional activity was determined by ERE-luciferase (3X ERE TATA luc; 11354; Addgene, Watertown, MA, USA) [24 (link)]. The pCMV-Green Renilla Luc vector (pCMV-Ren; 16153; Thermo Fisher Scientific, Waltham, MA, USA) was used for normalization. The ratio of ARR3tk-eGFP/SV40-mCherry to pCMV-Ren and 3X ERE TATA luc to pCMV-Ren was 100:1. Dual-Luciferase^® Reporter Assay System (E1910; Promega, Madison, WI, USA) was used, and the signal was captured and recorded by microplate reader Infinite F200 (Tecan, Seestrasse, Switzerland).

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Tsoi H., Shi L., Leung M.H., Man E.P., So Z.Q., Chan W.L, & Khoo U.S. (2022). Overexpression of BQ323636.1 Modulated AR/IL-8/CXCR1 Axis to Confer Tamoxifen Resistance in ER-Positive Breast Cancer. Life, 12(1), 93.

Publication 2022

3X ERE TATA luc Dual-Luciferase® Reporter Assay System Infinite F200

Assay Luciferase Promega Renilla Sv40 Transcriptional Vector

Corresponding Organization : University of Hong Kong

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Variable analysis

independent variables

AR transcriptional activity
ER transcriptional activity

dependent variables

Luciferase activity (ARR3tk-eGFP/SV40-mCherry)
Luciferase activity (3X ERE TATA luc)

control variables

Ratio of ARR3tk-eGFP/SV40-mCherry to pCMV-Ren (100:1)
Ratio of 3X ERE TATA luc to pCMV-Ren (100:1)
Dual-Luciferase® Reporter Assay System
Microplate reader Infinite F200

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