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. 2026 Mar 28;12:121. doi: 10.1038/s41531-026-01319-6

Fig. 8. Modulation of O-GlcNAcylation levels regulates microglial differentiation toward M1 or M2 phenotypes.

Fig. 8

Primary murine microglial cells were treated with various concentrations of O-GlcNAc modulators to assess their effects on microglial polarization phenotypes. A Cells were treated with GlcN at concentrations of 0.1, 1, 5, and 10 mM for 24 h. Whole-cell lysates were analyzed by western blot using antibodies against CD86 (M1 marker), CD163, and Arg1 (M2 markers), and β-actin as a loading control. Densitometric quantification was performed and normalized to β-actin (n = 3/group). B Cells were treated with Thiamet G (ThiaG) at concentrations of 0.05, 0.1, 0.5, and 1 μM for 24 h. Western blot analyses and quantification were conducted as described in (A) (n = 3/group). C Cells were treated with OSMI-1 at concentrations of 0.1, 1, 10, and 20 μM for 24 h. Expression of CD86, CD163, and Arg1 was assessed via western blot and quantified relative to β-actin (n = 3/group). D To evaluate the effects of O-GlcNAc modulation compared to cytokine-induced polarization, cells were treated with IL-4 (20 ng/mL), Thiamet G (1 μM), or OSMI-1 (20 μM) for 24 h. Western blotting was performed for CD86, CD163, and Arg1 (n = 3/group). Data are presented as mean SEM; *p < 0.05, **p < 0.01, ***p < 0.001 versus control, #p < 0.05, ###p < 0.001 versus IL-4. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc multiple comparison test.