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

Fig. 1. Reduced O-GlcNAcylation accompanies neuroinflammation and pro-inflammatory microglial activation in the SN of PD patients.

Fig. 1

A Western blot analysis of midbrain tissue lysates was performed using TH and β-actin antibodies. Quantification of TH expression was normalized to β-actin (n = 3 per group). B Representative immunohistochemical images of TH in human midbrain sections. Scale bar = 50 μm (n = 3 per group). C Co-immunostaining of α-synuclein with TH-positive neurons in the human midbrain. Representative merged images are shown (n = 3 per group). D Immunostaining of α-synuclein in midbrain sections. Scale bar = 50 μm (n = 3 per group). E Western blot analysis of O-GlcNAcylation levels and the expression of OGT, OGA, GFAT2 and β-actin in midbrain lysates. Quantification was normalized to β-actin (n = 3 per group). F Representative immunofluorescent images of primary microglial cells stained for O-GlcNAc (green), Iba1 (red), and DAPI (blue). Scale bar = 10 μm (n = 3 per group). G Representative immunofluorescent images of primary microglial cells stained for ASC (green), Iba1 (red), and DAPI (blue). Scale bar = 10 μm (n = 3 per group). H Western blot analysis of NF-κB signaling components in midbrain lysates, including phospho-p65 (p-p65), total p65, phospho-IκB (p-IκB), total IκB, and β-actin. Quantification of p-p65 was normalized to p65; p-IκB was normalized to IκB (n = 3 per group). I Western blot analysis of neuroinflammatory markers in midbrain lysates, including TLR4, CD86, CD68, CD163, Arg1, and β-actin. Quantification was normalized to β-actin (n = 3 per group). J Western blot analysis of inflammasome-related proteins in midbrain lysates, including NLRP3, ASC, cleaved caspase-1, total caspase-1, and β-actin. Quantification of NLRP3 and ASC was normalized to β-actin; cleaved caspase-1 was normalized to total caspase-1 (n = 3 per group). K Western blot analysis of phosphorylated and total STAT3 and ERK in midbrain lysates. Quantification of p-STAT3 was normalized to STAT3; p-ERK was normalized to ERK (n = 3 per group). L Western blot analysis of PPARγ, phosphorylated CREB (p-CREB), total CREB, and β-actin in midbrain lysates. Quantification of p-CREB was normalized to total CREB; PPARγ was normalized to β-actin (n = 3 per group). Data are presented as mean SEM; *p < 0.05, **p < 0.01, ***p < 0.001. Statistical analysis was performed using Student t test.