Journal: Antioxidants

Article Title: Cross-Species Insights into In Vitro Maturation Defects of the Oocyte and Identification of Crucial Regulators for Sheep Oocyte Maturation

doi: 10.3390/antiox14121499

Figure Lengend Snippet: EFNA1 alleviates excessive misfolded protein accumulation in oocytes during IVM. ( A ) GSEA analysis of differential expression genes related to oxidative phosphorylation, glutathione metabolism, and reactive oxygen species in in vitro matured oocytes compared to in vivo . ( B ) Effect of 10 ng/mL EFNA1 on ROS and GSH levels in matured sheep oocytes. Intracellular ROS and GSH levels were measured using CM-H2DCFDA and CellTracker™ Blue CMF2HC fluorescent probes, respectively (Scale bar = 200 µm). ( C ) Effect of 10 ng/mL EFNA1 on protein aggregates content in matured sheep oocytes. The protein aggregates content was determined by proteoStat aggresome detection reagent labeling (Scale bar = 50 µm). ( D ) Effect of 10 ng/mL EFNA1 on proteasome activity in matured sheep oocytes. The proteasome activity was evaluated by Me4BodipyFL (Scale bar = 50 µm). ( E , F ) Immunofluorescence detection of ER stress-associated protein (GRP78, ATF6) in matured sheep oocytes cultured with or without EFNA1 (Scale bar = 50 µm). ( G ) Relative expression levels of ER stress-associated genes (GRP78, PERK, EIF2A, ATF4, ATF6, IRE1, XBP1) in matured sheep oocytes cultured with or without EFNA1 as analyzed by RT-PCR. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: ROS and GSH levels were evaluated using the ROS assay kit (S0033S, Beyotime, Jiangsu, China) and CMF2HC (HY-D1571, MedChemexpress, Princeton, NJ, USA).

Techniques: Quantitative Proteomics, Phospho-proteomics, In Vitro, In Vivo, Labeling, Activity Assay, Immunofluorescence, Cell Culture, Expressing, Reverse Transcription Polymerase Chain Reaction