Research Article
O-GlcNAcylation Is Essential for Autophagy in Cardiomyocytes
Figure 3
OGT regulates the early stage of autophagy in cardiomyocytes. (a) Left panel, western blot analysis for autophagy of untreated or Bafilomycin A-treated Ad-LacZ or Ad-Cre-infected Ogtf/f neonatal cardiomyocytes, using antibody against O-GlcNAc, OGT, and LC3. GAPDH was detected as loading control. Right panel, quantification of western blot results in left panel, for each group. Significantly different; M.W.: molecular weight. (b) Left panel, western blot analysis for autophagy of Ad-LacZ- or Ad-Cre-infected Ogtf/f and/or Ogtf/y neonatal cardiomyocytes with either full medium (nutrient) or starving medium (starved), using antibody against pATG16L. GAPDH was detected as a loading control. Right panel, quantification of western blot results in the left panel, for each group. Significantly different; M.W.: molecular weight. (c) Left panel, representative immunomicroscopic images of pATG16L puncta (green) in Ad-LacZ- and Ad-Cre-infected Ogtf/f and/or Ogtf/y neonatal cardiomyocytes, costained with alpha-actinin (gray) and DAPI (blue). Right panel, quantification of pATG16L-positive puncta per cell, cells each group. Significantly different.
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