Skip to main content
. Author manuscript; available in PMC: 2013 Oct 1.
Published in final edited form as: Nat Chem Biol. 2013 Feb 24;9(4):271–276. doi: 10.1038/nchembio.1185

Figure 3. Compound (2) disrupts HIF-2 heterodimerization in vitro.

Figure 3

(a) Addition of (2) blocks heterodimer assembly between purified recombinant HIF-2α PAS-B* and ARNT PAS-B* heterodimer (squares) as assessed in the AlphaScreen Assay. No effect was observed in control reactions employing a single (doubly-tagged) GST-ARNT-PAS-B*-FLAG protein capable of recruiting both beads to induce an AlphaScreen signal (circles). Assays were performed in triplicate and the error bars represent ± SD. RU = relative units. (b) Compound (2) disrupts heterodimerization of the full length HIF-2 transcription factor. Nuclear extracts prepared from hypoxic Hep3B cells expressing ARNT, HIF-1α and HIF-2α (input) were incubated with increasing concentrations of (2). Immunoblot analysis indicates amounts of HIF polypeptides immunoprecipitated in the absence (-Ab) or presence of an anti-ARNT antibody.