Determination of ADAM17 gene promoter activity
Wild type and mutant GRE ADAM17 promoter (-150 to +200bp) were cloned
into pGL3-luciferase reporter vector (GenePharma Corporation). Primary
trophoblasts were transfected with the pGL3-luciferase reporter DNA and
pRL-TK-Renilla-luciferase plasmid (Promega Corp., Beijing, China) using
LipofectamineTM 3000 for 12h
(He et al., 2014). Cells were then
treated with DEX (10-6M) for 24h. Luciferase assays
were carried out using the dual luciferase assay kit (Promega).