Luciferase Assay for BCL2 Promoter

The luciferase reporter assay was performed as described previously [24 (link)]. The BCL2 promoter was cloned into the pGL3-Basic luciferase plasmid (Promega, USA) to construct the WT BCL2 reporter. BCL2 truncat plasmids and mutant plasmids were also constructed (Fig. 5E, H). For the Renilla luciferase reporter assay, each reporter construct was co-transfected into HEK293T cells together with the KLF5 plasmid or control plasmid. After 48 h of incubation, luciferase activity was measured using the Dual Luciferase Reporter Assay System (Promega, USA).

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Shen X., Zhang Y., Xu Z., Gao H., Feng W., Li W., Miao Y., Xu Z., Zong Y., Zhao J, & Lu A. (2022). KLF5 inhibition overcomes oxaliplatin resistance in patient-derived colorectal cancer organoids by restoring apoptotic response. Cell Death & Disease, 13(4), 303.

Publication 2022

pGL3-Basic luciferase plasmid Dual Luciferase Reporter Assay System

Assay Bcl2 Cells Luciferase Pgl3 Plasmid Promega Renilla luciferase

Corresponding Organization : Ruijin Hospital

Other organizations : Shanghai Institute of Hematology

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Variable analysis

independent variables

BCL2 promoter constructs (WT BCL2 reporter, BCL2 truncated plasmids, BCL2 mutant plasmids)

dependent variables

Luciferase activity

control variables

Renilla luciferase reporter
Control plasmid

controls

Positive control: WT BCL2 reporter
Negative control: Control plasmid

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