Research Paper Volume 11, Issue 4 pp 1110—1128

The ubiquitin ligase KBTBD8 regulates PKM1 levels via Erk1/2 and Aurora A to ensure oocyte quality

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Figure 4. KBTBD8 is essential for meiosis and regulates multiple key kinases. A. Western blot confirmation of KBTBD8 depletion by specific antibody transfection in oocytes. GAPDH was used as a loading control. B. Quantification of KBTBD8 levels in control and KBTBD8-depleted oocytes. C. KBTBD8 depletion dramatically disrupted spindle organization in MI oocytes. MTs: microtubules; Kinets: kinetochores. D. Representative bright-field images of control and KBTBD8-depleted oocytes. Numbers in the image indicate ratio of 1PB oocytes/total oocytes. E. Quantification of 1PB extrusion rate. F. Western blots showing decreased p-Akt and cyclin B1, and increased p-Cdk1 expression in KBTBD8-depleted oocytes. Tubulin was used as a loading control. G. Co-IP results demonstrating that KBTBD8 interacts with cul3. Scale bars, 20 µm for C, 200 µm for D. *p ˂ 0.05.