Fig. 4

Knocking down IP3R1 prevents the metabolic decline in oocytes from obese mice. a The mRNA levels of Itpr1 after knocking down IP3R1. b The relative mRNA levels of Itpr1. N = 90 per group across three replicates. c Representative immunoblots showing IP3R1 protein levels after IP3R1 knockdown. d The relative protein levels of IP3R1. N = 300 per group across three replicates. e Characteristic TEM images of oocytes from NC, siIP3R1 and siPACS-2 treatment. Magnification, 60,000×; scale bars, 500 nm. f The length of the ER tethered to the mitochondria was normalized to the ER length. N = 30 (five sections per mice, six mice per group) for each group across three replicates. g [Ca²⁺]_m levels after IP3R1 knockdown; scale bars, 50 μm. h Quantitative analysis of [Ca²⁺]_m. N = 30 per group across three replicates. i Representative immunoblots of increased P-MAPK 3/1 and decreased cytochrome C levels after IP3R1 or PACS-2 knockdown (left column, NC siRNAs; middle column, Itpr1 pooled siRNAs; right column, Pacs-2 pooled siRNAs). j Quantification of the immunoblots. N = 300 per group across three replicates. k ROS and GSH levels after IP3R1 knockdown; scale bars, 100 μm. l Statistical analysis of ROS and GSH levels after IP3R1 knockdown. N = 30 per group across three replicates. Graphs show means ± SEM. * P < 0.05, ** P < 0.01, CD vs. HFD; t-test