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. Author manuscript; available in PMC: 2018 Aug 18.
Published in final edited form as: ACS Chem Biol. 2017 Jun 19;12(8):2030–2039. doi: 10.1021/acschembio.7b00232

Figure 4.

Figure 4

Thiamet-G increases global O-GlcNAcylation in differentiating hESCs but does not affect differentiation kinetics. (A) Western blot shows that O-GlcNAc levels increase with Thiamet-G treatment from day –2 to 11. (B) Western blot shows that treatment with Thiamet-G does not affect differentiation kinetics since no difference in OCT4 and PAX6 protein expression was observed in Thiamet-G-treated cells in comparison to DMSO-treated cells. (C) Immunofluorescence analysis of TUJ1 expression levels in differentiated hESCs treated with DMSO, Thiamet-G, or Ac4-5SGlcNAc from day –2 to 11. Thiamet-G treatment does not recapitulate the neuronal phenotype obtained in Ac4-5SGlcNAc-treated hESCs. (D) Percent coexpression of TUJ1 and MAP2 in cells treated with DMSO, Thiamet-G, or Ac4-5SGlcNAc for the entire duration of neural differentiation, as determined by flow cytometry analysis, confirms the results obtained by immunofluorescence (n = 6; mean ± SEM; ***P < 0.001).