Figure 6.

Pin1 inhibits transcription initiation in vitro. (A) Linear CMV promoter-containing DNA templates were preincubated with NE for 10 min at 30°C, then pulse-labeled with [α-³²P]UTP for 30 sec at room temperature (lane 1) or supplemented with cold NTPs and incubated for 10 min at 30°C (lane 2). (B) The same DNA templates were preincubated with NE as in A but pulse-labeled without (lane 1) or with 100 and 200 ng of wild-type (lanes 2,3) or R68,69A mutant (lanes 4,5) Pin1. (C) Linearized DNA templates were preincubated and pulse-labeled as in B, but with cold NTPs only (lane 1) or plus 200 ng of wild-type (lane 2) or R68,69A mutant (lane 3) Pin1 and then incubated for 5 min at 30°C. (D) Similar pulse experiment as B but with mock-depleted (lane 1), or Pin1-depleted (lane 2) NEs as in Figure 5, or Pin1-depleted NE plus wild-type (lane 3) or R68,69A mutant (lane 4) Pin1. (E) Quantitative analyses of data from three independent experiments as in B (top) and D (bottom).