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. 2012 Jan 1;3(1):101–110. doi: 10.4161/nucl.18955

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Copyright © 2012 Landes Bioscience

This is an open-access article licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. The article may be redistributed, reproduced, and reused for non-commercial purposes, provided the original source is properly cited.

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graphic file with name nucl-3-101-g2.jpg — Figure 2. F3H interaction screen for M18BP1 interaction partners. (A) Scheme depicting the F3H screening strategy. Cells containing a lac operator array were transfected with plasmids expressing a lac repressor-GBP fusion protein, M18BP1-EGFP and mCherry/RFP-CCAN proteins. The lac repressor binds to the lac operator array and through the GBP recruits M18BP1-EGFP. CCAN proteins interacting with M18BP1 are consequently enriched at the lac operator array. (B) Representative examples for M18BP1 interacting (CENP-C) and non-interacting (CENP-A and CENP-W) proteins are shown. Scale bar is 5µm. (C) Summary of interaction tests between M18BP1 and CCAN proteins. Interactions were tested with the F3H assay using M18BP1-EGFP and 16 RFP or mCherry fusions with CCAN proteins. From all 16 tested CCAN proteins, only CENP-C showed a clear interaction with M18BP1.