Fig. 1
From: O-GlcNAc regulation of autophagy and α-synuclein homeostasis; implications for Parkinson’s disease
Inhibition of OGA by thiamet G (TG) increased O-GlcNAcylated protein levels in neurons. Primary cortical neurons were cultured from E18 rat embryos. At DIV7, neurons were exposed to TG (0.25, 2.5 and 25 μM) for 24 h (a) and 7 d (b). For detection of O-GlcNAc modified proteins, 10 μg protein was separated on a 7% SDS-PAGE and subjected to immunoblotting using CTD110.6 monoclonal antibody against O-GlcNAcylated proteins. β-actin was used as a loading control. (c-d) Quantification of the band intensities for O-GlcNAc modified proteins from (a-b) assessed with image J software 1.48e NIH, USA. (e-f) Cell viability was measured by the trypan blue exclusion method. For all panels, Data = mean ± SEM (n = 3), normalized to 0 μM TG. *p < 0.05 compared to 0 μM TG control, by ANOVA followed by Bonferroni’s Multiple Comparison post-hoc test. The exact p value is given above each bar for this comparison. The levels of protein O-GlcNAcylation among the 3 thiamet G concentrations were not significantly different