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. 2011 May 3;23(5):1756–1771. doi: 10.1105/tpc.111.084590

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© 2011 American Society of Plant Biologists

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Figure 5. — BAF1 Proteins Form Homo- and Heterodimers.

(A) Schematic representation of the constructs used in yeast two-hybrid and in vitro pull-down assays.

(B) Targeted yeast two-hybrid assay. Yeast transformants were grown on selective medium for both bait (BD:BAF1-Δ762) and prey (AD:BAF1, AD:BAF1R-1, and AD) constructs (-LT) and on medium lacking His (-LTH) for detecting interactions. Interactions are also verified by β-galactosidase activity (βgal). AD, GAL4 activation domain; BD, GAL4 DNA binding domain.

(C) and (D) In vitro pull-down assays. BAF1 (C) and BAF1R-1 (D) fused to a 3XHA tag were obtained by in vitro transcription/translation and were incubated with GST-only (negative control) or GST-BAF1 bound beads. Anti-GST antibody shows even loading of GST-only and GST-BAF1 proteins, whereas anti-HA antibody shows preferential retention of the input only on GST-BAF1 beads.