Luciferase reporter assay

To investigate whether FOXC2 is transcriptionally regulated by p53, PANC-1 cells were transfected with FOXC2 promoter (P2/P3-WT or P2/P3-MUT) reporter constructs, and further co-transfected with sip53. After a 48-hour culture, the Dual-Luciferase reporter assay system was used to assess promoter activity, with Renilla luciferase used for normalization.

To determine how HIF-1α and HDAC1 affect the activity of HRE on the CF129 promoter under hypoxia condition. PANC-1 and BxPC-1 cells expressing pGL3-based construct containing the HRE of CF129 were transfected with siNC and siHIF-1α, and further treated with and without TSA. Twenty-four hours later, cells were cultured with or without CoCl2 for an additional 24h. The reporter activity was measured as above.