Identification of effective Gli1 binding sites on the S100A4 promoter.

<p>A: S100A4 luciferase activity increases with increasing expression level of SHh in AsPC-1 cells. The pGL3-1.5 S100A4 and Renilla luciferase vectors were transiently transfected into L-Gli1i infected, L-C infected or L-Shh infected AsPC-1 cells respectively. Results were normalized for transfection efficiency using Renilla luciferase and the L-C infected group was arbitrarily given a value of 1. B: Relative luciferase activity of the S100A4 promoter Gli1 binding site mutants. AsPC-1 cells transfected by L-Shh were transiently transfected 5mg of each reporter construct including pGL3-1.5 S100A4, pGL3-1.5 S100A4 Mut1, Mut2 and Mut3 respectively. The pGL3-1.5 S100A4 was arbitrarily given a value of 1 and the activities of the other transfections were adjusted relative to this activity. C: Site 1 was responsible for Gli1 transcription. Relative luciferase activity from different AsPC-1 cell groups, L-Gli1i infection, L-Shh infection or L-C infection, transfected by different constructs, pGL3 1.5 S100A4 Mut 1–2, Mut 2–3 and Mut 1–3 with Renilla luciferase vectors. Results were normalized for transfection efficiency using Renilla luciferase and the L-C infected group was arbitrarily given a value of 1. * <i>P</i><0.05, ** <i>P</i><0.01.