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Fig. S4
Related to Figure 2.
(A) Time-lapse of a neuroepithelial cell in a Tg(HS-OS-Plk4) fish undergoing a bipolar division. H2B-RFP labels chromatin and Ras-mKate2 labels cell outline (both green), centrin-GFP labels centrosomes (magenta). The yellow arrows mark the position of amplified centrosomes. Nuclei are highlighted with dots. HS applied at 24hpf, imaging started at 12hphs. The frames are from Movie S1. Scale bar is 5µm. Time is in min.
(B)-(C) En face confocal scans of mitotic neuroepithelial cells in the Tg(OS-Plk4) fish, HS applied at 24hpf, fixed at 16hphs, featuring bipolar (B) and multipolar (C) anaphase with unbalanced centrin composition at the poles, visualized by centrin antibody staining (magenta). Merged with cell outlines (phalloidin,blue) and chromatin (DAPI, green). Scale bar is 5µm.
(D) Distribution of mitotic lengths divided into time subcategories (upper three graphs) and lengths of mitosis depicted on the single cell level (lower three graphs) of neuroepithelial cells. Mitotic length was measured from the time the chromatin starts condensing to the time daughter nuclei are furthermost apart in the anaphase. Left graphs: mitotic length of all cells in wr fish, wr fish injected with HS-OS-Plk4 and Tg(HS-OS-Plk4) fish. Middle graphs:mitotic lengths of cells in wr fish injected with HS-OS-Plk4 shown in two groups based on anaphase type (bipolar or multipolar). Right graphs: mitotic lengths of cells in Tg(HS-OS-Plk4) fish shown in two groups based on anaphase type (bipolar or multipolar). On all three lower graphs bars represent mean± SEM. n marks cell number, N=13 (WT fish), N=20 (injected OS-Plk4 fish), N=10 (transgenic OS-Plk4 fish)