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. 2015 Jun 2;6:7162. doi: 10.1038/ncomms8162

Figure 4. A region encompassing CD1 in Bub1 is required for RZZ localization.

Figure 4

(a) Schematic of Bub1 primary structure with motifs indicated and below a schematic of the different truncation constructs used in this study. All constructs contain the Bub3 binding site ensuring kinetochore targeting. (b) HeLa cells were depleted of endogenous Bub1 using RNAi and the indicated Bub1 constructs were co-transfected for 48 h and cells were arrested with nocodazole for 2 h. Cells were fixed and stained for Bub1, CREST and ZW10. (c,d) The Bub1 (c) or ZW10 (d) kinetochores levels was measured and normalized to CREST signals. At least 160 single kinetochores were analysed from eight different cells and the mean with standard error of mean is indicated. A t-test was used to compare the values in d. (NS, not significant (P>0.05), *P≤0.05, **P≤0.01, ****P≤0.0001). Scale bar, 5 μm.