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

Fig. 5. GlcN and Thiamet G modulates nuclear translocation of NF-κB subunits p65 and c-Rel in microglia.

Fig. 5

LPS (5 μg) was injected into the SN region of mouse brains. GlcN (200 mg/kg) or Thiamet G (ThiaG; 20 mg/kg) was administered intraperitoneally three times per week for 4 weeks (A–D). A, C Cytoplasmic (CE) and nuclear (NE) protein fractions were isolated from SN tissue. Western blot analysis was performed using antibodies against NF-κB subunits p65 and c-Rel. Nuclear levels of p65 and c-Rel were normalized to their corresponding cytoplasmic levels. H2B and α-tubulin were used as nuclear and cytoplasmic markers, respectively (n = 3/group). Wheat germ agglutinin (WGA) pull-down assays were performed using total SN protein lysates to assess O-GlcNAcylation of p65 and c-Rel. O-GlcNAc-modified forms were detected by western blot and normalized to total p65 or c-Rel levels (n = 3/group). E–H Primary microglial cells were treated with LPS (400 ng/mL) for 24 h in the presence or absence of GlcN (5 mM) or Thiamet G (ThiaG; 1 μM). E, G Nuclear and cytoplasmic fractions were analyzed for p65 and c-Rel distribution by western blot, with quantification normalized as described above (n = 3/group). F, H WGA pull-down assays were conducted on total cell lysates to assess O-GlcNAcylation of p65 and c-Rel (n = 3/group). Data are presented as mean SEM; *p < 0.05, **p < 0.01, ***p < 0.001 versus control, #p < 0.05, ##p < 0.01, ###p < 0.001 versus LPS. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc multiple comparison test.