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. 2010 Oct 12;123(21):3789–3795. doi: 10.1242/jcs.073387

Fig. 2.

Fig. 2.

CLP-1 associates with HDACs at the onset of C2C12 cell differentiation. (A) Immunoprecipitation (IP) of C2C12 cell lysates from growth (G) and differentiation medium at 24 hours (D1) and at 72 hours (D3) with antibodies specific for HDAC1, HDAC3 and HDAC5, and western blot with anti-CLP-1 and anti-cdk9 antibodies. IP with antibody alone served as a control for immunoreactivity (Ab). Western blot with anti-CLP-1 and anti-GAPDH antibodies served as input controls. Data represent one of three separate experiments for each antibody. Bottom right panel: western blot of total cell lysates with anti-HDAC1, anti-HDAC3 and anti-HDAC5 antibodies. Western blot with GAPDH served as input control. (B) Left panel: nuclear (Nuc) and cytoplasmic (Cyto) fractions of C2C12 cells in differentiation medium at 24 hours were subjected to direct western blotting with antibodies specific for CLP-1, cyclin T1, MyoD and GAPDH. Right panel: lysates were subjected to IP with anti-HDAC5 antibody and western blotting with anti-CLP-1 antibody. Direct western blot of CLP-1 served as an input control.