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OPP assays were performed as described previously (30). Two specificity controls were included: (i) cells not labelled with OPP and (ii) cells incubated with cycloheximide (#C7698, Sigma) at a final concentration of 50 μg/ml for 15 min prior to, as well as during incubation with OPP.
For quantitation of OPP labeling, cells were analysed by flow cytometry. Selection of whole single cells in G1 phase was based on forward and side scatter, as well as on DAPI staining. Analysis of fluorescence intensities was performed in Kaluza 1.2 (Beckman Coulter).
Copyright and License information: ©2019 The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
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