Journal:

Article Title: Regulation of Subnuclear Localization Is Associated with a Mechanism for Nuclear Receptor Corepression by RIP140

doi: 10.1128/MCB.23.12.4187-4198.2003

Figure Lengend Snippet: Interaction between endogenous GR and RIP140. (A) RIP140 was coimmunoprecipitated with GR from HTC cells with anti-GR antibody-Sepharose (anti-GR) in the absence (−) or presence (+) of the glucocorticoid ligand TA. Coimmunoprecipitation with normal mouse antibody-Sepharose (control ab) was used as a negative control. Proteins in the cell extract used for the precipitation (input) and precipitated proteins (ppt) were analyzed by Western blotting with anti-RIP140 and anti-GR antibodies. IP, immunoprecipitate. (B) In vitro-translated RIP140 or RIP140 domains were incubated with GST or the GST-GR LBD bound to glutathione beads. The interacting proteins were visualized by SDS-PAGE and autoradiography. The values on the left are molecular sizes in kilodaltons.

Article Snippet: The Sepharose pellet was washed with EPG buffer containing 50 mM NaCl three times, and bound proteins were eluted by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer and analyzed by SDS-PAGE and Western blotting with anti-RIP140 polyclonal antibody (H-300; Santa Cruz Biotechnology) and anti-GR monoclonal antibody (250; Ann-Charlotte Wikström, Karolinska Institutet).

Techniques: Negative Control, Western Blot, In Vitro, Incubation, SDS Page, Autoradiography