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. 2023 Jun 23;299(8):104950. doi: 10.1016/j.jbc.2023.104950

Figure 7.

Figure 7

Favoring O-GlcNAcylation by O-GlcNAcase (OGA) inhibition increases abnormal spindles in defolliculated oocytes (DEs).Xenopus DEs and folliculated oocytes (FCs) expressing epidermal growth factor receptor (EGFR) were microinjected with Grb7 (15 ng) or not, incubated or not with 100 μM OGA inhibitor Thiamet G (ThG), 1 h before epidermal growth factor (EGF) stimulation (5 nM). DEs were submitted to 500 nM of integrin inhibitor echistatin (Echist). A and B, the normal, abnormal, or absent spindles were scored in oocytes fixed, dehydrated, and paraffin-embedded on sections (7 μm) stained with nuclear red (nuclei and chromosome detection) and picroindigocarmine (cytoplasmic structure detection). C, Grb7-GST and EGFR immunoprecipitations were realized and followed by Western blotting with anti-O-GlcNAcylation (O-GlcNAc), anti-GST (Grb7-GST), and anti-OGT antibodies. D, sWGA pulldowns were performed before Western blotting with anti-O-GlcNAcylation (O-GlcNAc) and anti-OGT antibodies. Results were obtained on 10 to 20 oocytes from three different females. Statistical significance was accepted for ∗∗p < 0.01, ∗∗∗p < 0.005, and ∗∗∗∗p < 0.001. GST, glutathione-S-transferase; sWGA, succinylated wheat germ agglutinin.