Fig. 4.

REV-ERBα impacts on protein O-GlcNAcylation in both cytoplasmic and nuclear compartments. (A) Label-free mass spectrometry analysis of O-GlcNAcylated proteins. Control (IgG) or O-GlcNAcylated protein-enriched (RL2) immunoprecipitates from siRNA (Scr or REV-ERBα)-transfected HepG2 cells were analyzed by label-free mass spectrometry analysis. (B) LC-MS/MS analysis of O-GlcNAcylated proteins. HepG2 cells were treated like in A, and immunoprecipitates were fractionated by SDS/PAGE before mass spectrometry analysis. (C) Cellular abundance of AKT and H2B in siRNA-transfected HepG2 cells. HepG2 cells were cultured at 25 mM Glc. Cyt, cytoplasm; ER, endoplasmic reticulum; Mb, plasma membrane; Mt, mitochondria; Nuc, nucleoplasm.