Fig 2: Confirmation glycoprotein of interest is MAP6. (A) Western blot analysis of soluble lysates and insoluble precipitates. M, protein size standards. (B) Densitometric analysis of MAP6 levels normalized to the level of β-actin in soluble lysates and insoluble precipitates. Bars depict the mean±SD; * p<0.05 vs. control. (C) Blots showing that MAP6 is a glycoprotein containing Gal-(β-1,3)-GalNAc oligosacchrides. Immunoprecipitation was coupled with Western blot analysis of extract from the whole mouse brain. The SDS-PAGE gel was visualized using silver-staining kit; and the membrane of WB was blotted with ABL, and visualized using a ECL kit.

Fig 3: Dual immunofluorescence analysis of MAP6 glycosylation in brainstems from MPTP-induced mice. Photomicrographs show co-localization of MAP6 (A; red) and ABL (B; green). Nuclei are stained with DAPI (blue). Merged images show their colocalization (MAP6+ABL) (C and D; yellow). White arrows point to MAP6 with a glycoprotein. The enlarged image is the boxed part in the merged image.

Fig 4: In situ dual immunofluorescence analysis glycosylated MAP6 in various brain areas in PD mice. (A) Representative photomicrographs of immunofluorescent labeling of glycoproteins detected by ABL (green) and MAP6 stained using anti-MAP6 monoclonal antibody (red) in the indicated areas, as well as their colocalization (yellow). Nuclei were stained using DAPI (blue). (B) Statistical comparison of the immunofluorescent signals between the MPTP-induced PD mice and control mice. Bars depict the mean±SD; *p<0.05 vs. control group.