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. 2012 Feb 15;287(15):11602–11615. doi: 10.1074/jbc.M112.351601

FIGURE 1.

FIGURE 1.

Cdo interacts with ASK1 and TAK1. A, lysates of 293T cells transiently transfected with the GST-Cdo intracellular region (GST-Cdointra), FLAG-tagged ASK1 (ASK1(FLAG)), or control (−) expression vectors, as indicated, were pulled down with glutathione-Sepharose beads and blotted (WB, Western blot) with antibodies to FLAG or the Cdo intracellular region. Total lysates were also immunoblotted with the indicated antibodies. B, lysates of 293T cells transiently transfected with GST-Cdo intracellular region, HA-tagged TAK1 (TAK1(HA)), or control (−) expression vectors, as indicated, were pulled down with glutathione-Sepharose beads and blotted with antibodies to HA or the Cdo intracellular region. Total lysates were also immunoblotted with the indicated antibodies. C, lysates of 293T cells transiently transfected with Cdo, ASK1(FLAG), S epitope-tagged JLP (JLP(S)), or control (−) expression vectors, as indicated, were pulled down with S-agarose and immunoblotted with antibodies to Cdo, FLAG, or S-epitope. Total lysates were also immunoblotted with the indicated antibodies. D, lysates of 293T cells transiently transfected with Cdo, TAK1(HA), JLP(S), or control (−) expression vectors, as indicated, were pulled down with S-agarose and immunoblotted with antibodies to Cdo, HA, or S-epitope. Total lysates were immunoblotted with the indicated antibodies. E, lysates of C2C12 cells that were proliferating in growth medium (lane G), at near confluence (day 0) or in DM for the indicated times were immunoprecipitated with anti-Cdo antibody and immunoblotted with anti-ASK1, anti-TAK1, anti-JLP, or anti-Cdo antibodies. Total lysates were also immunoblotted with antibodies to ASK1, TAK1, JLP, Cdo, and MHC (an indicator of differentiation) and to cadherin as a loading control. F, quantification of three independent experiments shown in E. The intensity of the ASK1 and TAK1 signals was quantified, and the values from condition lane G were set to 1.0. Values represent the means of triplicate determinations ± 1 S.D.