2.5. Reporter assay
RGM1 cells were seeded at a density of 7.0 × 104/well in a 24- well plate and cultured for 24 h to reach sub-confluence. Reporter constructs containing different length Cox-2 promoters, a gift from Dr. H. Inoue (National Cardiovascular Center Research Institute, Osaka, Japan), are shown in Fig. 1. The
Fig. 1. Schematic representation of reporter vectors containing the 5′-flanking region of the Cox-2 gene promoter. The construct of long-type of reporter vector containing the 5′-flanking region of the COX-2 gene promoter contains 3 promoter sites (NF-IL6, NF-κB, and CRE) (−327/+59). No promoter is contained in the short-type of reporter vector containing the 5′-flanking region of the COX-2 gene promoter (−52/+59). Distances are given as nucleotide positions relative to the transcriptional start site as +1.