A) 2 kb region directly upstream of the translation initiation site (ATG codon) was cloned into a firefly luciferase expression vector, pGL3, by PCR (3’ primer is overlined on top of the sequence). Transcription start sites in the DBTSS (http://dbtss.hgc.jp) database are indicated with right-angled arrows below the sequence, and by numbers of full-length cDNAs representing a particular TSS. A schematic representation of the cloned region is shown with predicted binding sites for SMAD transcription factors (CAGACA, underlined), for the transcription factor KLF2 (core binding site CACCC, italicized sequences) and binding site for MYC/MAX dimers (CACGTG, underlined). Nucleotide position numbers represent the numbers used by DBTSS. B) The promoter construct, with or without the SV40 enhancer, and in forward and reverse orientations, were transfected into 293T cells. Mean relative promoter activities (normalized by Renilla activity) 48 h post transfection are shown from duplicate transfections +/− S.D.