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. 2022 Aug 15;119(34):e2202821119. doi: 10.1073/pnas.2202821119

Fig. 5.

Fig. 5.

O-GlcNAc of Gli2 S355 suppresses Gli2 acetylation by inhibiting binding to HAT. (A and B) Representative immunoblot of AcGli2 and O-GlcNAc of Gli2 S355 (Gli2 S355 O-GlcNAc) wtih (A) WT OGT or (B) H498A/H558A double mutant OGT (OGTMut), respectively. (C) Representative immunoblot of AcGli2 and Gli2 S355 O-GlcNAc with Myc-tagged Gli2. Transfected cells were treated with OSMI-1 (10 μM) or TM-G (80 μM) for 36 h. (D) Representative immunoblot of AcGli2 and Gli2 S355 O-GlcNAc with Myc-tagged Gli2 or Myc-tagged Gli2 S355A (Myc-Gli2 S355A), respectively. (E) Representative immunoblot of AcGli2 and Gli2 S355 O-GlcNAc with Myc-tagged Gli2 or Myc-Gli2 S355A, treated with OSMI-1 (10 μM) or TM-G (80 μM) for 36 h, respectively. (F) Representative immunoblot AcGli2 and Gli2 S355 O-GlcNAc immunoprecipitated from NIH 3T3 cells transfected with Myc-tagged Gli2 in the presence of SAG (0.1 μM) for 36 h, followed by OSMI-1 (10 μM) or TM-G (80 μM) treatment for 24 h. (G) Representative immunoblot of HA-p300 with Myc-tagged Gli2, with or without GFP-tagged OGT. (H) Representative immunoblot of HA-p300 with Myc-tagged Gli2. Transfected cells were treated with OSMI-1 (10 μM) or TM-G (80 μM) for 36 h. (I) Representative immunoblot of HA-p300 with of Myc-tagged Gli2, or Myc-Gli2 S355A mutant, respectively. See also SI Appendix, Fig. S12.