Conventional G-less cassette transcription assay.
(A) Micrococcal nuclease (MNase) digestion of chromatin-assembled p208p21ML plasmid. 500 ng of assembled chromatin was digested with MNase and resolved using 1.2% agarose gel electrophoresis and ethidium bromide staining. The mass ratios of core histones to DNA are indicated. 100 bp DNA ladders were used as a size marker. (B) Summary of the in vitro transcription reaction is outlined (left to right) from chromatin assembly to the transcription reaction involving [α-32P] UTP-mediated labelling. PIC, preinitiation complex. (C) p53- and p300-dependent transcription of naked (left panel) and chromatin-assembled (right panel) p208p21ML plasmid. G-less cassette transcript of 365 bp was resolved in the 8 M urea-PAGE gel and visualized by autoradiography. Relative signal intensities were quantified using phosphoimager software and values are indicated at the bottom of the autoradiogram.
