By Aria Baniahmad (auth.), Aria Baniahmad (eds.)
Because thyroid hormone receptors (TRs) are vital regulators of a giant variety of organic processes-ranging from mind improvement to manage of imaginative and prescient in vertebrates-abnormal functioning of those receptors could have severe clinical results. In Thyroid Hormone Receptors: tools and Protocols, a panel of remarkable investigators surveys and explains the main state of the art tools utilized in TR learn and their functional experimental info. defined in step by step element to make sure strong experimental effects, the options offered conceal a wide selection of key components, together with TRs in improvement and iteration of knockout (mouse and Xenopus), transcriptional legislation via TRs in either cell-free structures and dwelling cells, and TR mutant research of sufferers. extra tools supply robust instruments for the isolation of TR-regulated protein complexes, for the research of blood phone differentiation throughout the oncogene v-erbA, and for objective gene research within the mind. Microarray chip tools also are provided for studying the organs of transgenic mice to spot goal genes within the liver. every one protocol has been constructed via a hands-on professional and comprises complete explanatory notes, troubleshooting information, and important reviews on fending off pitfalls.
extensive ranging and hugely useful, Thyroid Hormone Receptors: tools and Protocols deals endocrinologists, geneticists, biochemists, and learn physicians a finished choice of robust equipment necessary not just for all significant parts of TR biology and research, but additionally for nuclear hormone receptors in general.
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Additional info for Thyroid Hormone Receptors: Methods and Protocols
Tsai, S. , O’Malley, B. , and Tsai, M. J. (1997) Gene silencing by chicken ovalbumin upstream promoter-transcription factor I (COUP-TFI) is mediated by transcriptional corepressors, nuclear receptor-corepressor (N-CoR) and silencing mediator for retinoic acid receptor and thyroid hormone receptor (SMRT). Mol. Endocrinol. 11, 714–724. 19. , et al. (1999) The nuclear receptor corepressor N-CoR regulates differentiation: N-CoR directly interacts with MyoD. Mol. Endocrinol. 13, 1155–1168. 20. , et al.
4. 5 M EDTA (see Note 4). 5. Add 10 µL 1 N NaOH and incubate at 65°C for 20 min to hydrolyze residual RNA. Cool to room temperature (see Note 5). 6. 5. 7. 5, and 20 µg of human Cot-1 DNA to a MicroCon 30 cartridge. Pipet to mix and spin for 10 min at 16,000g. 8. 5, and concentrating to about 20–30 µL (approx 5 min at 16,000g). 9. Recover by inverting the concentrator over a clean collection tube and spinning for 2 min at 16,000g (see Note 6). 10. Take a 2–3 µL aliquot of the Cy5-labeled cDNA for analysis, leaving 18–28 µL for hybridization.
And Fig. 1. Similar results were obtained in a repeat analysis with different mice from the same lines. (49–51). Myc promotes cell proliferation, transformation, or apoptosis, requires its dimerization with Max, and binds to its cognate DNA recognization E-box site (CACGTG) (53). It was shown that Mad/Max/Sin3/NCoR/HDAC complexes could repress transcription and potentially inhibit growth (7–10). Thus, Mad/Max and Mxi1/Max dimers can act as antagonists of Myc binding to Myc enhancer sites and repress transcription by recruiting corepressors (54–55).