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Fig. 3. Loss of Tum results in mislocalisation of other cytokinetic furrow components in mutant embryos. (A,B) The localisation of AurB (green) is shown alone in the upper panels, and merged with the microtubule (red) and DNA (blue) images in the lower panels in wild-type embryos (A) and tum^DH15 embryos (B). AurB is associated with the chromosomes of metaphase cells (arrowheads, A and B) in mutant and wild-type embryos. At anaphase AurB moves to the central spindle and cortex in wild-type cells (arrow, A) but is not localised correctly in mutant cells (arrow, B). (C,D) The localisation of Diaphanous (green) is shown alone in the upper panels, and merged with the microtubule (red) and DNA (blue) images in the lower panels in wild-type (C) and tum^DH15 (D) embryos. Diaphanous is cortical throughout the cell cycle, but concentrates at the midzone cortex of wild-type telophase cells (arrowheads, C). In tum^DH15 mutants, Diaphanous does not concentrate in the midzone cortex (arrowhead, D). (E,F,G) The localisation of Pbl-GFP (green) is shown alone in the upper panels and merged with the microtubule (red E, G), Tum (red, F) and DNA (blue) images in the lower panels in wild-type (E,F) and tum^DH15 (G) embryos. At anaphase, Pbl-GFP is concentrated in the midzone cortex in normal cells (arrowheads, E,F) but remains cortical throughout mitosis in tum^DH15 mutant cells (arrowheads, G). (H,I) The localisation of Anillin (green) is shown alone in the upper panels, and merged with Tum (red) and DNA (blue) in the lower panels in wild-type (H) and tum^DH15 (I) embryos. Anillin concentrates at the midzone at anaphase (arrowhead, H) and concentrates in the furrow as cells undergo cytokinesis (arrow, H). In tum mutant cells, Anillin is cortical in metaphase (asterisk, I) and does not concentrate in the midzone cortex during telophase (arrow, I). (J,K) The localisation of Pav-KLP (green) is shown alone in the upper panels, and merged with the microtubule (red) and DNA (blue) images in the lower panels in wild type (J) and tum^DH15 (K) embryos. In wild-type embryos, Pav-KLP concentrates at the furrow at anaphase (arrow, J) and becomes concentrated in the midbody as cytokinesis progresses (arrowhead, J). In tum mutant embryos, no Pav-KLP is detected in anaphase or telophase cells (arrowhead, K). Bar, 5 µm. Grayscale images for all color channels can be viewed in supplementary material Fig. S3.

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