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. 2004 Feb 12;101(8):2339–2344. doi: 10.1073/pnas.0308676100

Fig. 1.

Fig. 1.

ARC92 is a VP16-targeted subunit of the ARC/Mediator coactivator complex. (A) ARC92 was purified as a VP16-interacting protein. HeLa cell nuclear proteins were first bound to the phosphocellulose (PC) column and were then successively eluted by the indicated concentration of KCl. The PC fractions were incubated with GST-VP16 beads, and specifically bound proteins were visualized by SDS/PAGE and silver stain. The band indicated by the arrowhead was identified as human ARC92 by peptide microsequencing. (B) Schematic representation of ARC92 and the related PTOV1 proteins. The amino acid sequence of one portion in ARC92 (gray box) is highly similar to two segments of PTOV1, a protein overexpressed in prostate tumors. (C) The ARC/Mediator complex is coimmunoprecipitated with the overexpressed Flag-ARC92. Flag-tagged ARC92 was expressed in U2OS cells and immunoprecipitated from nuclear extract by anti-Flag or anti-HA (IP control) antibodies and analyzed by immunoblotting for the indicated proteins. (D) Endogenous ARC92 is coimmunoprecipitated with the ARC/Mediator complex. Anti-TRAP220/ARC205 and anti-Med6 specifically coimmunoprecipitated endogenous ARC92 along with other components of the ARC/Mediator complexes from HeLa cell nuclear extracts. Other nuclear proteins, such as p300 and PTOV1, were not coprecipitated.