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. Author manuscript; available in PMC: 2009 Aug 10.
Published in final edited form as: Nature. 2008 May 7;453(7198):1132–1136. doi: 10.1038/nature06923

Figure 1. A FRET-based sensor of Aurora B kinase activity demonstrates a spatial phosphorylation gradient during anaphase.

Figure 1

a, Sensor design: phosphorylated threonine underlined, targeting sequences from histone H2B (chromatin) or CENP-B (centromere). b, Cells expressing cytosolic (untargeted), chromatin-targeted, or centromere-targeted sensors were imaged live through anaphase. The YFP/CFP emission ratio at each timepoint was normalized to vary from 0–100% and averaged over multiple cells (N≥4). c–e, Cells expressing the centromere-targeted sensor were imaged through anaphase, either with an intact checkpoint (c) or with Mad2 depleted by RNAi (d–e). Left panels (c,d): unprocessed YFP images, right panels: color-coded images of the emission ratio, timestamps (min) relative to anaphase onset, scale bars 5 um. In a plot of all timepoints (e), each circle represents an individual centromere characterized by time after anaphase onset, position along the division axis, and emission ratio (color scale). Dashed lines indicate data points plotted in Fig. S4.