DNA replication progression was analyzed using the DNA fiber assay (Xu et al., 2011 (link); Liu et al., 2016 (link)). Cells were labeled with IdU (25 μM) for 30 min, followed by exposure to HU (4 mM) or DMSO for 4 h, and chased with CldU (125 μM) for 40 min. Cells were lysed, DNA fibers were spread, and the IdU and CldU tracts were stained and detected, as previously described (Xu et al., 2011 (link); Liu et al., 2016 (link)). DNA fibers were imaged using a Leica DM4000 fluorescent microscope.
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DNA Replication Progression Assay
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Corresponding Organization : City Of Hope National Medical Center
Other organizations : Nanjing Normal University
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Variable analysis
independent variables
- Exposure to HU (4 mM) or DMSO for 4 h
- DNA replication progression
- Cells were labeled with IdU (25 μM) for 30 min
- Cells were chased with CldU (125 μM) for 40 min
- Cells were lysed, DNA fibers were spread, and the IdU and CldU tracts were stained and detected
- DMSO treatment as a negative control
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