Huh7 cells were transfected at 70% confluency in DMEM growth media containing 5% FBS with 8xGTIIC-Tead4-luciferase (Addgene), a 624b YAP1 enhancer-luciferase construct with or without the mutations in the first intronic TCF site7 (link), and renilla-luciferase plasmids (Addgene) using Biolab transfection reagent. After overnight, the cells were treated with 12(S)-HHTrE (50 nM, cat#34590),12-HETE (100 nM, cat#34570), Ozagrel (1 µM, cat#70515), LY255283 (10 µM, cat#70715) obtained from Cayman Chemical Company and the YAP-TEAD binding inhibitor Super-TDU (1-31) TFA (300 nM, MedChemexpress, cat#HY-P1728A) with respective vehicle controls in serum-free medium for 24 h. Protein concentrations were determined in cell lysates and equal amounts of proteins were analyzed for firefly and renilla luciferase activities using Dual-Glo Luciferase Assay Kit (Promega).
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