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. 2020 Aug 4;8:764. doi: 10.3389/fcell.2020.00764

FIGURE 3.

FIGURE 3

Nitroblue tetrazolium (NBT) staining for assessing the effect of superoxide ion scavenger (Tempol) on intracellular ROS in oocytes and cumulus cells. Ovine cumulus-oocyte complexes (COC) were matured in vitro without (control) or with Tempol (1, 10, 25, 50, or 100 mM) supplementation and stained with NBT. The dark purple/blue spots of formazan precipitates represent ROS. The level of intracellular ROS (% NBT stained area, mean ± SE) was determined in oocytes and cumulus cells from five individual COC in each experimental group. (A) Unstained COC, control group; (B) NBT-stained COC, control group; (C–G) NBT-stained COC, Tempol supplemented groups; (H,I) level of intracellular ROS in oocytes and cumulus cells in different experimental groups. Values without a common superscript (a,b,c,d) above error bar differ significantly (P < 0.05). BF, bright-field image; PH, phase-contrast image. Scale bar = 100 μm.