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. Author manuscript; available in PMC: 2013 Mar 1.
Published in final edited form as: Mol Cancer Res. 2012 Jan 24;10(3):378–391. doi: 10.1158/1541-7786.MCR-11-0314

Figure 2. LEDGF/p75 interacts with MeCP2 in vitro.

Figure 2

(A) Pull down assays with His-LEDGF/p75 and GST-MeCP2. Recombinant His-LEDGF/p75 was incubated with GST or GST-MeCP2 bound to glutathione beads, and pulled down proteins were analyzed by immunoblotting using antibodies specific for GST or LEDGF/p75. His-FADD (Fas-associated protein with death domain), was used as negative control. Protein input was determined by immunoblotting of whole cell extracts. *Denotes degraded GST-MeCP2. (B) Cross titration for Flag-LEDGF/p75 and GST-MeCP2 interaction as measured by AlphaScreen® assay. Interaction was measured at different concentrations of Flag-LEDGF/p75 as indicated on the vertical legend, and GST-MeCP2 as indicated on the X-axis. (C) An estimated 1 nM MeCP2 was sufficient to interact with 11 nM LEDGF/p75. E.coli BL21 lysate not expressing GST-MeCP2 was used as a negative control. Results in (B) and (C) are representative of three independent measurements.