Modulation of O-GlcNAcylation Regulates Autophagy in Cortical Astrocytes

Figure 1

Pharmacological inhibition of OGA and OGT modulates autophagy in cortical astrocytes. (a) Immunoblot analysis of OGA, OGT, and O-GlcNAcylation levels in astrocytes treated with thiamet-G (1 μM) or alloxan (5 mM) for 24 h. (b) Dot blot analysis of O-GlcNAcylation levels after thiamet-G or alloxan treatment for 24 h. Densitometry quantification of O-GlcNAcylation protein was performed by ImageJ. All statistical analyses were obtained as (, ). (c) Autophagic flux in astrocytes was visualized by immunofluorescence staining with anti-GFAP (red) and anti-LC3 (green) antibodies using confocal microscopy. (d) LC3 puncta formation was measured from at least 3 independent random areas from each slide, and the puncta from a minimum of 5 cells which contained them were counted and calculated (ns = nonsignificant, ). (e) Immunoblot analysis of LC3 and p62 levels in astrocytes treated with thiamet-G or alloxan (, ns = nonsignificant, , ).

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