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Cells were grown on glass coverslips in 24-well trays, and after 24 h cells were treated as required then incubated with 25 μM IdU for 20 min. Cells were then fixed in 3% buffered paraformaldehyde in PBS for 10 min at room temperature, and DNA denatured via a 30 min incubation with 2.5M HCl. Indirect immunofluorescence was performed using an anti-mouse BrdU antibody (BD, clone 3D4, 1:250) that cross-reacts with IdU, an Alexa anti-mouse 488 secondary antibody (Invitrogen, 1:1000) and DAPI counterstaining.
Copyright and License information: ©2024 The Author(s) 2024. 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 (https://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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