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. 2005 Aug 1;19(15):1755–1760. doi: 10.1101/gad.347005

Figure 2.

Figure 2.

PHO and PCC bind synergistically to the bxd PRE. (A) PRE binding of PHO, ΔPBD and PCC was studied by bandshift assays. Binding reactions contained ∼5 nM PHO and/or ∼5-15 nM PCC, and radiolabeled PHO4/5-PRE. PRE binding of PHO and PCC was analyzed by primer extension DNaseI footprinting using either naked DNA (B,C) or chromatinized PHO4/5-PRE (D) templates. Binding reactions on naked DNA contained ∼20 nM (+) or 100 nM (‡) PHO and ∼20-60 nM PCC. Chromatin binding reactions contained ∼50 nM PHO and ∼20-60 nM PCC. DNaseI digestion ladders were analyzed by primer extension, resolved on a 6% polyacrylamide gel, and visualized by autoradiography. The positions of the PHO sites and footprinted areas are indicated. Closed arrowheads indicate DNaseI hypersensitive sites induced by protein binding.