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. 2008 Jul 7;28(17):5458–5468. doi: 10.1128/MCB.00549-08

FIG. 2.

FIG. 2.

The 5′ enhancer, the promoter region, and the 3′ control region are active in pancreatic acinar cell lines, but only the 5′ enhancer is responsive to PTF1. (A) The 5′ enhancer together with the promoter region, the promoter region alone, or the 3′ control region was linked to a minimal promoter (mp) and a luciferase (Luc) reporter gene (see Materials and Methods), and AR4-2J pancreatic acinar cells or 293 nonpancreatic cells were transfected with the constructs. (B) To determine which regions were responsive to the PTF1 complex, 293 cells were cotransfected with the same constructs and expression plasmids for the three components of the PTF1 complex. +, present. (C) To test whether the activity of the enhancer required the PTF1 binding sites, AR4-2J and 266-6 acinar cells and 293 nonacinar cells were transfected with the 5′ enhancer linked to a minimal promoter driving luciferase expression or with the same construct bearing mutations in the proximal, the distal, or both PTF1 binding sites. (D) To verify that the PTF1 complex acted through the known PTF1 binding sites, 293 cells were cotransfected with the 5′ enhancer construct or the 5′ enhancer construct with both TC boxes mutated and plasmids expressing components of the PTF1 trimer. Transfections were normalized according to the expression levels of a pCMVβ plasmid with which the cells were cotransfected. Error bars represent standard deviations.

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