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. 2010 Jan 28;29(6):1105–1115. doi: 10.1038/emboj.2010.3

Figure 2.

Figure 2

Differential interaction of SWI/SNF and Mediator complexes with R3 methylated and unmethylated LAP*/C/EBPβ1 N-terminal peptides. (A) The N-terminal sequence of LAP*/C/EBPβ1 was synthesized with R3 in unmodified form (1–41) or in asymmetrically dimethylated form (1–41 Rme2a). Peptides with covalently attached C-terminal biotin moieties (grey pentagons) were applied to UNIPEX proteomic libraries and interactions were revealed by streptavidin-HRP (SA-HRP) and ECL. Positive clones were identified as characteristic duplicate pattern per square. Libraries contained 48x48 squares with four duplicate clones on two PVDF membranes. A single square containing the brama-related gene 1 (Brg1) expression clone interacts with unmethylated, but not with methylated peptide (arrow heads). (B) Unmethylated and methylated C/EBPβ peptides (1 μM), as shown in (A) were incubated with cell lysates and bound proteins separated by streptavidin Dynabeads. Cell lysates were prepared from Raji cells (top) or HEK-293 MED23-HA-transfected cell lysates (bottom). Western blots were incubated with anti-hBrm, anti-BAF155, anti-BAF47/Ini1, anti-HA, and anti-MED26 as indicated.