The gene promoter sequences of TGFβ1, from -3515 to -2501 base pair containing 4 Klf2 binding sites were inserted into a pGL3 promoter luciferase vector. The primer sequences for amplification of Klf2 binding sequence are listed in Table S5 and the final sequence was validated by DNA sequencing. Luciferase activity was determined after transfection for 48 h using Dual-Luciferase assay kits (Promega, Madison, WI, USA). Individual luciferase activity was normalized to the corresponding renilla-luciferase activity, and the data presented are the average of five luciferase assays performed in triplicate.

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