Figure 5
From: Hepatoma derived growth factor binds DNA through the N-terminal PWWP domain
HDGF represses the transcription of a reporter which is controlled by the HDGF DNA binding element. The 80 bp oligo (Hcis) was cloned upstream of the SV40 promoter which controls a downstream luciferase reporter gene. 0.5 ug Hcis-SV40-LUC reporter construct (A) or SV40-LUC control reporter (B) were cotransfected with various amount (0.1, 0.3, 0.5 ug) of GFP-HDGF. 0.1 ug pRL-CMV were included as transfection control. The dual-luciferase reporter assay system was used to measure the luciferase activities 24 hours after transfection. Percentage of firefly luciferase activity was calculated when luciferase activity from the reporter gene alone was set at 100%. Firefly luciferase activity was normalized with constitutive renilla luciferase activity to correct for transfection efficiency. Bars indicate mean ± SEM from 3 separate transfection assays with duplicate plates, *P < 0.05 by one-way ANOVA control versus GFP-HDGF.