FIG. 2.

Effect of OGT inhibition using BADGP on levels of embryonic O-GlcNAcylation assessed at 68 h post-hCG. Representative confocal immunofluorescent images of embryos immunolabeled with RL2 antiserum following culture in KSOM (0.2 mM glucose; A) or KSOM supplemented with OGT inhibitor BADGP at 1 and 2 mM (B and D, respectively). Immunoreactivity of RL2 in the presence of DMSO (solvent control) was also assessed (C). Original magnification ×250; bar = 25 μm. The mean gray scale intensity of at least two nuclei in the same optical plane for each embryo was quantified using Image J and averaged to yield one value per embryo. For each treatment in each of two experiments, at least five embryos were analyzed, and data are presented graphically as mean nuclear O-GlcNAc immunoreactivity ± SEM. Factorial ANOVA revealed no interexperimental variation or interaction and a significant decrease in RL2 immunoreactivity in response to 2 mM BADGP (Tukey post hoc test, ***P < 0.001).