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. 2018 May 30;7(7):bio028225. doi: 10.1242/bio.028225

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© 2018. Published by The Company of Biologists Ltd

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 use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 1. — Cif-mediated CRL substrate stabilisation. (A,B) HEK293T cells were transfected with FLAG-tagged Cif, V5-tagged Cif or Cif C156S for 48 h and treated with 1 μM MLN4924 for the last 24 h. Cell lysates were resolved using SDS-PAGE and the cellular abundance of CRL substrates was analysed using Western blotting. (C) Cells were transfected with V5-tagged Cif_Bp or Cif_Yp for 48 h before cell lysis. (D,E) Cells were transfected with V5-tagged Cif_Bp or Cif_Yp for 48 h before chase with 40 μM cycloheximide for the indicated times, followed by cell lysis and Western blotting.