Fig. 6: Metaphase I spindle and chromosome-microtubule attachment in fixed oocytes after 10 h culture and cold treatment. | Nature Communications

Fig. 6: Metaphase I spindle and chromosome-microtubule attachment in fixed oocytes after 10 h culture and cold treatment.

From: CENP-V is required for proper chromosome segregation through interaction with spindle microtubules in mouse oocytes

Fig. 6

a Percentage of cells showing chromosome misalignment. Chromosome misalignment is defined when the chromosome is entirely outside the region of the equatorial plate (transparent area in c) (b) Average number of chromosome misalignments per oocyte. (c) Example of Cenp-V+/+ and Cenp-V−/− oocytes. Microtubules (left) and DNA (center) are stained by anti ß-tubulin and Hoechst 33258 respectively. See also supplementary Movies S6 and S7. Scale bar = 10 µm. (d) Spindle pole width (SPP width; w, dashed line in c) in Cenp-V++ and Cenp-V−/− oocytes. (e) K-fiber quantification expressed as arbitrary units (A.U.) of fluorescence within the boxed region shown in c. (f) Confocal planes of chromosome-microtubule attachments in Cenp-V+/+ and Cenp-V−/− oocytes. A maximum projection of the whole spindle is shown for each genotype. A proper attachment to two sister-centromeres is indicated by a white dashed square in Cenp-V+/+. For Cenp-V−/− oocytes different types of attachments are shown: two sister centromere attachment (white); attachment to one sister centromere (pink); and unattached sister centromeres (yellow). The tip of the K-fiber attached to the centromere is indicated by a yellow arrowhead. Centromeres are identified as the strongest region stained by Hoechst 33258. Please see also supplementary Movies S8 and S9. Scale bar = 10 µm. (g) Quantification of chromosome-microtubule attachment. Statistic: data were expressed as mean ± SD and statistical differences were tested by a paired two tailed T- test (***p < 0.001, **p < 0.01, *p < 0.05). The exact p values are available in the source data file. Statistical differences refer to the average of all data points. (d, e) Data are represented as box plots where the middle line is the median, the lower and upper boxes correspond to the first and third quartiles and outlying points are plotted beyond the end of the whiskers. (a, b) n = 30 cells; (d) n = 21 cells, (e) n = 24 cells and (g) n = 113 attachments. All data come from 3 independent experiments.

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