Figure 3

Determination of the promoter activity of the intergenic region. The directions of gene transcription are indicated by arrows. Here, a 1053-bp fragment (from position -718 to +335) consisting of the entire 405-bp intergenic region and ~300 bp beyond its two boundaries is cloned as the full-length promoter construct. Progressive deletions from this full-length promoter construct were produced and inserted into the promoterless vector in either the sense (relative to PREPL) or antisense (relative to C2ORF34) direction. The resulting luciferase activity of transiently transfected U87MG cells is given adjacent to each construct. Co-transfection of pRL-TK plasmid was performed for normalization of transfection efficiencies and cell viability. Units are relative to SV40 control promoter, defined as 1×. The experiments were performed in triplicate. Data are representative of three independent experiments, and error bars are ± standard deviation. A similar result was also observed in human H4 cells (data not shown).