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. Author manuscript; available in PMC: 2014 Sep 5.
Published in final edited form as: Cell. 2014 May 22;157(5):1061–1072. doi: 10.1016/j.cell.2014.03.046

Figure 2. TRPM7 binds transcription factors and subunits of chromatin remodeling complexes.

Figure 2

A. Endogenous TRPM7 was IP’d from combined cytosolic+nuclear extracts of WT3 or KO9 mESC with αA25 mouse or αC47 rabbit antibody and the IP probed on WB with antibody to the indicated proteins. INO80 cell lysate lanes were run on a different gel, as indicated by gap. B. Subunits of Polycomb and INO80 chromatin remodeling complexes bind ectopically expressed M7CKs. FLAG-tagged transcription factors were transiently co-expressed in 293T cells with HA-tagged M7CK-L, -M or -S. Cytosolic+nuclear extract was IP’d with αHA-agarose and co-IP’d proteins were probed on WB with αFLAG-peroxidase. C. Affinity-purified long form of M7CK (M7CK-L) binds purified transcription factors (Coomassie). Left 3 panels: HA-tagged M7CK-L (M7) was combined with purified YY1 or Ezh2 protein complexes and IP’d with αHA-agarose. Right panel: purified GST or GST-RYBP fragment (M7CK-binding domain, a. a. 22–47, GST-RYBPbd) fusion proteins were combined with purified M7CK-L and GST pulled down with glutathione agarose.