Cell cycle arrest can be performed just by analysing the intensity and condensation of DNA since the intensity of the Hoechst dye depends on the DNA content. Mitosis cells (4n DNA content) have a doubled dye intensity when compared to G1 cells (2n DNA content). This information can be used to classifie the cells between the different phases:
RO-3306, a selective CDK1 inhibitor, stops reversibly proliferating human cells at the G2/M phase. This effect can be observed through Hoechst dye analysis:
Other dyes can be used to identifie the cell cycle arrest more specifically. Here is an example of mitosis arrest by colchicine, observed through histone H3 phosphorylation:
Phospho-histone H3 can allow us to look at the different steps of the cell mitosis phases:
With colchicine, cells have been arrested at the end of G2 phasis / prophasis:
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