BrdU incorporation assay was used to determine the effects of nicotine on cell proliferation. This assay was performed using a BrdU incorporation assay kit (Cell Signaling, Danvers, Massachusetts) as described previously.21 (link) Briefly, 48 hours after nicotine treatment, mESCs were treated with 10 mM BrdU for 4 hours and incubated with anti-BrdU peroxidase conjugate for 90 minutes before adding a tetramethylbenzidine substrate. Incorporated BrdU was measured by an enzyme-linked immunosorbent assay reader, and data were represented as fold change compared to control group.
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Nicotine Impacts on Cell Proliferation
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Corresponding Organization : Binzhou Medical University
Other organizations : Affiliated Hospital of Qingdao University, Qingdao University, Yuhuangding Hospital, Shandong University, Jinan Central Hospital
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Variable analysis
independent variables
- Nicotine treatment
- Cell proliferation (measured by BrdU incorporation)
- Time of BrdU treatment (4 hours)
- Incubation time with anti-BrdU peroxidase conjugate (90 minutes)
- Control group (not treated with nicotine)
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