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 1. PKM 1 is essential for oocyte maturation and quality. A. Western blot results confirming nearly complete PKM1 depletion induced by specific antibody transfection. GAPDH was used as a loading control. B. Bright field images of control and PKM1-depleted (PKM1-DE) oocytes. Arrowheads indicate oocytes containing first polar bodies (1PB). Numbers indicate the ratio of 1PB oocytes/total oocytes. C. Quantification of 1PB presence in control and PKM1-depleted oocytes. D. ATP quantification in control and PKM1-depleted oocytes. E. JC1 fluorescence indicative of decreased mitochondrial membrane potential after PKM1 depletion. JC1 aggregates are red, monomers are green. F. Quantification of JC1 aggregate/monomer ratio in control and PKM1-depleted oocytes. G. ROS generation, revealed by DCFH-DA staining (green fluorescence), in control and PKM1-depleted oocytes. BF: Bright field. H. Quantification of ROS levels in control and PKM1-depleted oocytes. Scale bar for B, 200 µm; for E and G, 20 µm. * p ˂ 0.05.