Figure 4.

Hyperacetylation of tubulin‐K40 impairs kinetochore–microtubule attachments during oocyte meiosis. (a) WT, WT + K40R, WT + K40Q oocytes were stained with α‐tubulin antibody to visualize spindle (green) and counterstained with PI to visualize chromosome (red). Arrows point to the misaligned chromosomes and arrowheads indicate the disorganized spindle. Scale bars: 20 µm. (b) Quantification of WT, WT + K40R, WT + K40Q oocytes with spindle/chromosome defects. (c) WT, K40Q and K40R mutant cRNA were microinjected into fully grown oocytes for analysis. Metaphase oocytes were stained with CREST for kinetochores (purple), anti‐tubulin antibody for microtubules (green), and Hoechst 33,342 for chromosome (blue). Representative confocal images are shown. Scale bars: 5 µm. (d) Quantitative analysis of K‐MT misattachments in WT (n = 16), K40R (n = 18), and K40Q (n = 13) oocytes. Attachment of kinetochores to microtubules was assessed through examination of the full series of Z‐axis focal planes. Kinetochores in regions where fibers were not easily visualized were not included in the analysis. Scale bars: 20 µm. *p < .05 versus controls