Skip to main content
. Author manuscript; available in PMC: 2016 May 1.
Published in final edited form as: Gene. 2015 Feb 19;561(2):235–248. doi: 10.1016/j.gene.2015.02.050

Figure 9. Methylation of the SLC9B1 CpGII does not affect transcriptional elongation efficiency in HEK 293 cells.

Figure 9

(A) Schematic representation of the tandem reporter construct (TAN1) construct, which consists of two quantifiable reporter vectors (firefly luciferase and Renilla luciferase). In between the two reporters is a linker region, which has a multiple cloning site for insertion of the test sequence (in this case CpGII). Flanking the test sequence are self-cleaving ribozyme sites, which ensure that the test sequence does not become part of the either of the reporter mRNA fragments. Transcription initiates at the human CMV promoter, proceeds through the firefly luciferase, through the test sequence and then the Renilla luciferase ultimately resulting in the generation of both the firefly and Renilla proteins. The ratio of Renilla luciferase to firefly luciferase activity provides a measure of the transcription elongation efficiency. (B) The effect of in vitro methylation of SLC9B1 CpGII on transcriptional elongation efficiency was determined. The dual luciferase reporter assay was used to determine the Renilla/firefly activity ratio in HEK 293 transfected with the mock methylated (MM) or in vitro methylated (IV) CpGII in the TAN1 reporter construct. The Renilla/firefly activity ratio of the TAN1-NHEDC1-CpGII IV was normalized to the TAN1-NHEDC1-CpGII-MM (set at 1.0). The data is represented as mean±SD of six independent transfections.