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. 2014 Oct 18;66(1):85–97. doi: 10.1093/jxb/eru401

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© The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 6. — Split-ubiquitin assay used to detect PPIs among XyG biosynthetic proteins. Transformed yeast containing the indicated combinations of TF–Cub and NubG fused proteins were spotted in an OD₅₄₆ of 1.5 and up to 1000× dilution on SD-His-Leu and SD-His-Leu-Trp-Ade plates. Growth on SD-His-Leu-Trp-Ade plates indicates a positive interaction. X-Gal assay performed on growing yeast on SD-His-Leu is a test for β-galactosidase activity, a reporter for interaction upon blue colour formation, Ost1p–NubI (NubI) and pPR3-N test for the functionality and random interaction of the Cub-fused proteins, respectively. The type II membrane protein TF–Cub–Anp1p tests for random interaction among NubG-fused proteins. Consensus of three biological replicates is shown. (This figure is available in colour at JXB online.)