Figure7. Identification of compounds that induce EHF overexpression.

(A-B). PANC-1, BxPC-3 and two primary cancer cell lines PDX1# and PDX2# were treated with rosiglitazone(5μM and 10μM,24h). DMSO was used as control. (A) Q-PCR was conducted to detect for EHF mRNA expression. (B) Western-blot for EHF expression were performed. Representative results were shown. (C) PPAR γ-scanned motif logo (D) Predicted PPAR γ response elements (PPREs) on the human EHF promoter. Position relative to the transcription start site of EHF, PPRE sequences and corresponding JASPAR scores. (E) Binding of PPAR γ to the promoter of EHF was determined by chromatin immunoprecipitation. IgG was used as negative control. Anti-RNA PolymeraseⅡwas used as positive control. Representative results were shown. (F) The promoter activity of EHF after treated with rosiglitazone. PANC-1 transfected with either luciferase reporter pGL3-empty vector or wild type pGL3-ESE3/EHF promoter were treated with rosiglitazone(10μM,24h). Forty-eight hours later, cells were collected for dual luciferase assay. Results were expressed as fold induction relative to those of the corresponding cells transfected with pGL3-empty vector after normalization of firefly luciferase activity according to Renilla luciferase activity. All experiments were repeated three times independently. Paired Student’s t-test was used as statistical analysis. *P<0.05 and **P<0.01.