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. 2020 Apr 1;219(6):e201907082. doi: 10.1083/jcb.201907082

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© 2020 Jackman et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 6. — MAD1 remains associated with TPR and condensing chromosomes when it cannot bind cyclin B1. (A) Prometaphase parental RPE cyclin B1-Venus^+/−:Ruby-MAD2^+/− cells and the MAD1 E53/E56K clones 7D2 and 8B12 were fixed and stained with Hoechst 33342 (blue), anti-TPR (green), anti-MAD1 (red), and ACA antibodies as indicated. Scale bar, 2 µm. (B) Quantification of MAD1-TPR colocalization percentage shown in A (parental n = 12 cells; 7D2 n = 10 cells; 8B12 n = 10 cells). (C) Cells were fixed and stained as in A except that they were pretreated with 166 nM reversine. Scale bar, 2 µm. Images are representative of two independent experiments. (D) Quantification of MAD1-TPR colocalization percentage shown in C (parental n = 11 cells; 7D2 n = 11 cells; 8B12 n = 9 cells).