Research Paper Volume 13, Issue 23 pp 25393—25407

Figure 3. Risk allele A of rs34166160 exhibits a higher transcriptional activity through interacting with C/EBP beta. (A) Data from luciferase assays in Hela cells. Reporters containing the 1020 bp length genomic fragment overlapping rs34166160 harbored A allele or C allele were co-transfected with pENTER-C/EBP beta or a negative control pENTER into Hela cells. Cells were harvested 48 h after transfection and luciferase activities were measured and normalized to renilla activities. (B) Data from luciferase assays in Hela cells. Reporters containing a 30 bp length core genomic segment overlapping rs34166160 harboring the A allele or C allele were co-transfected with pENTER-C/EBP beta or a negative control pENTER into Hela cells. The measurement of luciferase activities was previously described. Three independent experiments were performed. Error bars represent standard deviation (SD). ^*P < 0.05. (C) The risk allele A of rs34166160 can directly bind to the transcription factor C/EBP beta. EMSA probe containing risk allele A or wide type allele C of rs34166160 incubated with (lanes 2–3; lane 5-6) or without (lane 1; lane 4) nuclear extracts from HUVECs transfected with pENTER-C/EBP beta. Lane 1 and lane 4, 5′ -end biotin-labeled probe alone; lane 2 and lane 5, EMSA for 5′ -end biotin-labeled probe and HUVECs nuclear extracts; lane3 and lane 6, excessive unlabelled probe and HUVECs nuclear extract.