Figure 4

Identification of the Jmjd6 – 1110005A03Rik bi-directional transcriptional unit. (A) Schematic overview of the 65 bp non-coding and non-overlapping intergenic region between the murine Jmjd6 and 1110005A03Rik (Rik) genes. The conserved sequence element identified using PipMaker in Figure 2 is shown in orange. 5'-untranslated regions (UTRs) are indicated in grey, and first exon coding sequences of the Jmjd6 gene in green and of the 1110005A03Rik gene in blue, respectively. The region indicated by two half arrows was used for promoter activity assays presented in (B). It was amplified and cloned to yield an 840 bp test fragment (B) This putative bi-directional promoter fragment was used in a luciferase reporter gene assay to measure transcriptional activity. Basal promoter activity was found for both transcriptional orientations compared to a promoterless control (pGL3) vector. The 5'-3' orientation of the fragment (1110005A03Rik orientation, Rik) yielded only half of the transcriptional activity as it was measured for the 3'-5' orientation (Jmjd6 orientation, Jmjd6). The values reported for the transfection experiments are means ± standard deviation of three independent triplicate experiments. P value was determined using Student's t-test. *** P < 0.001.