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. 2015 May 19;35(3):e00193. doi: 10.1042/BSR20150038

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© 2015 The Author(s) This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC-BY) (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.

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

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Cells were grown at 30°C to early log phase, then pulse-labelled with ³⁵S-methionine for 5 min. Following the addition of 20 mM cold methionine and 10 μg/ml cycloheximide, samples were incubated at 30°C or 37°C and then frozen in 10% TCA at the specified time points. Tim9 antibodies were used to immunoprecipitate Tim9 and labelled Tim9 was detected by autoradiography. Levels were quantified and expressed as a percentage of intensity at 0 min.