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. 2014 Jun 19;10(6):e1004415. doi: 10.1371/journal.pgen.1004415

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© 2014 Vanoosthuyse et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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A. Serial dilutions of the indicated strains were plated on rich media at the indicated temperatures. B. The CPF component Yth1 tagged at the endogenous locus with 3flag epitopes was immuno-precipitated from cycling cells in the presence or absence of Swd2.2. Whole cell extracts (WCE) and the immuno-precipitated material (Flag IP) were analyzed by western blot. Yth1-3flag interacts with Pfs2-GFP whether or not Swd2.2 is present. Arrows indicate aspecific bands on the western blot. C. Asynchronous populations of the indicated strains were grown at 30°C and ChIP-qPCR was performed to analyze the amount of Pfs2-GFP cross-linked to chromatin (mean ± standard deviation from 4 biological replicates). D. Western blot analysis of total protein extracts of the indicated strains. Tubulin (TAT1 antibody) is used as a loading control.