Figure 3.
Ash2L interacts preferentially with phosphorylated Mef2d and Mef2c. (a) Flag-tagged Mef2a, Mef2c or Mef2d treated with activated p38α (or untreated) were incubated with extract prepared from differentiating (48 h) C2C12 cells. After washing, Mef2-associated proteins were subjected to western blotting analysis with anti-Ash2L or anti-Flag. (b) Flag-tagged wild-type (WT) or mutant Mef2d were incubated with activated p38α and [32P]ATP. Reactions were separated by SDS-PAGE. (c) Interaction studies with Flag-tagged WT and mutant Mef2d as in a. (d) Cell extracts prepared from differentiating C2C12 cells (48 h) that had been incubated in the presence (+SB) or absence (−SB) of the p38 MAPK inhibitor SB203580 were subjected to immunoprecipitation with anti-Mef2 or control IgG. Immunoprecipitated proteins were analyzed by western blotting with indicated antibodies.