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

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.