HDAC Activity Is Required for Efficient Core Promoter Function at the Mouse Mammary Tumor Virus Promoter
Figure 4
Repression of MMTV transcription by TSA is consistently recapitulated in an in vitro transcription assay. (a) Flow chart for a typical experiment. Nuclear extracts are generated from Hela S3 cells which were either untreated or TSA treated. These extracts were used for in vitro transcription assays with the templates shown in (b). The MMTV template contains the region from the TATA box to the initiator element and a G-free cassette of 400 bp. The adenovirus major late (AdML) template contains a promoter fragment from −400 bp to +10 bp with a G-free cassette of 190 bp. Results of a typical transcription assay are shown in (c). C-extract from untreated cells (Control). T-extract from TSA-treated cells. (d) shows a graphic representation of assay results using three independent sets of extracts. Each extract was assayed with each template at least three times. Transcription levels in the extracts from TSA-treated cells are expressed relative to those in the corresponding extracts from untreated (control) cells.
