PRODH safeguards human naive pluripotency by limiting mitochondrial oxidative phosphorylation and reactive oxygen species production
Cheng Chen 1,2,8 , Qianyu Liu 3,8 , Wenjie Chen4,8 , Zhiyuan Gong5 , Bo Kang6 , Meihua Sui 2, Liming Huang 1 & Ying-Jie Wang 6,7
Abstract: Naive human embryonic stem cells (hESCs) that resemble the pre-implantation epiblasts are fueled by a combination of aerobic glycolysis and oxidative phosphorylation, but their mitochondrial regulators are poorly understood. Here we report that, proline dehydrogenase (PRODH), a mitochondria-localized proline metabolism enzyme, is dramatically upregulated in naive hESCs compared to their primed counterparts. The upregulation of PRODH is induced by a reduction in c-Myc expression that is dependent on PD0325901, a MEK inhibitor routinely present in naive hESC culture media. PRODH knockdown in naive hESCs significantly promoted mitochondrial oxidative phosphorylation (mtOXPHOS) and reactive oxygen species (ROS) production that triggered autophagy, DNA damage, and apoptosis. Remarkably, MitoQ, a mitochondria-targeted antioxidant, effectively restored the pluripotency and proliferation of PRODH-knockdown naive hESCs, indicating that PRODH maintains naive pluripotency by preventing excessive ROS production. Concomitantly, PRODH knockdown significantly slowed down the proteolytic degradation of multiple key mitochondrial electron transport chain complex proteins. Thus, we revealed a crucial role of PRODH in limiting mtOXPHOS and ROS production, and thereby safeguarding naive pluripotency of hESCs.
详情请见：https://doi.org/10.1038/s44319-024-00110-z
在该研究中圣尔生物通用型ECL发光液用于Western blotting实验
Western blotting (WB)
Finally, the blots were developed using the ECL reagent (Share-Bio, #sb-wb012) and visualized by the Tanon 5200 Multi Chemiluminescent Imager system.
