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. 2017 Oct 2;216(10):3117–3132. doi: 10.1083/jcb.201610012

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© 2017 Tipton et al.

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Figure 6. — Aurora B inhibition reduces Kif4A levels on mitotic chromosomes. (Left) Immunofluorescence of Kif4A, Kid, or histone H3 pS10 levels in metaphase HeLa cells treated with MG132 (MG) alone or in combination with ZM447439 (ZM) for 2 h to inhibit Aurora B activity. MG132 was included to prevent mitotic exit. Cells were labeled for DNA and kinetochores. Images represent maximum-intensity projections. Kif4A, Kid, and histone H3 pS10 levels are scaled equivalently in each panel. Bars, 10 µm. (Right) Fluorescence intensity quantification of chromosome-bound Kif4A, Kid, or histone H3 pS10 immunofluorescence after treatments. Loss of Aurora B kinase activity dramatically reduced Kif4A levels without affecting the levels of Kid. Loss of histone H3 pS10 levels was used to demonstrate inhibition of Aurora B kinase activity. SD is plotted for the mean from each condition. n = 20 cells from two independent experiments for each condition. ****, P < 0.0001. FI, fluorescence intensity.