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. 2014 May 26;205(4):541–554. doi: 10.1083/jcb.201307137

Figure 2.

Figure 2.

CENP-I increases the half-life of Mad1 at unattached kinetochores. (A) Images of Mad1-GFP FRAP in control, CENP-I–depleted, and ZM-treated cells arrested in nocodazole. (B) Recovery dynamics of Mad1-GFP after photobleaching demonstrating that CENP-I–depleted cells have a larger initial recovery of Mad1 and a faster turnover of stable Mad1. (C) Total recovery of Mad1-GFP at 120 s after photobleaching. (D) Scatter plot displaying the natural log of the normalized unrecovered fluorescence over time. The biphasic nature of Mad1 recovery is illustrated by overlaid lines. CENP-I–depleted cells have a fast phase of initial Mad1 recovery similar to controls but the pool of stable Mad1 in CENP-I–depleted cells has a greatly decreased half-life relative to control. Red arrows in A indicate FRAP targets. FRAP data are from n = 30 experiments. Error bars indicate standard deviation. *, P < 5 × 10−7; **, P < 2 × 10−11. Bars: (white) 5 µm; (yellow) 1 µm.