Heparin inhibits HGF-induced Egr1 expression and the Egr1 promoter activity in a dose-dependent manner.

HGF-induced activation of Egr1 mRNA and protein level (2A, 2C) and the effect of heparin on the HGF induced Egr1 expression in SK-HEP-1 cells were examined by RT-PCR and western blotting (2B, 2D). Egr1 transcriptional activity was measured by luciferase reporter assay after transient transfection of SK-HEP-1 with pGL2-Luc-B-Egr1 plasmid construct and the pRL-TK Renilia luciferase. Relative luciferase activity was determined as described in Materials and Methods. The firefly luciferase activity was normalized to Renilia luciferase activity (2E, 2F). Results are representative of two independent experiments performed in triplicate. Bars indicate standard error of the mean (SEM), asterisks (*) indicate statistically significant differences between the indicated groups.