AP1 activity assay

HC Haoming Chen
RP Ravi Padia
TL Tao Li
YL Yue Li
BL Bin Li
LJ Lingtao Jin
SH Shuang Huang
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AP1 transcriptional activity was measured with the aid of a firefly luciferase reporter gene plasmid (3xAP1pGL3) from Addgene (Watertown, MA). This plasmid contains three copies of AP1 consensus sequence and was previously used to monitor AP1 activity in B cells56. To determine cellular AP1 activity, this plasmid was co-transfected into cells with a Renilla luciferase gene-containing plasmid (pRL-TK, Promega, Madison, WI) using lipofectamin 2000 (Invitrogen, Carlsbad, CA). After 36 h, cells were lysed and cell lysates were assayed for luciferase activity using Dual-Luciferase® Reporter Assay System (Promega Corp, Madison, WI). Renella luciferase activity was used as an internal transfection control for standardization.

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