Fig. 2.

Jarid1a is tightly associated with the G9a-GLP heterodimer. (A) G9a, GLP, and Jarid1a coelute by size exclusion chromatography. A nuclear extract was separated on a Superose-6 column, and the presence of specific proteins in different fractions revealed by immunoblotting with Abs indicated on the left. Molecular masses (in kilodaltons) are indicated on the right. (B) G9a-GLP heterodimer associates with Jarid1a and the Mediator complex in a mutually exclusive manner. Proteins immunoprecipitated via Abs against Jarid1a and Med1 were analyzed by Western blot. Mock IPs with normal IgG were used as negative controls. Abs used for Western blot (Left) and molecular masses (Right; in kilodaltons) are indicated. (C) Jarid1a interacts directly with G9a. Rec purified Jarid1a or Ubc4 was incubated with rec purified G9a before IP with G9a Abs. Immunoprecipitated proteins were revealed by immunoblotting with the Abs indicated on the left.