Fig. 2.

GLN enhances global O-glycosylation pre-/post-heat shock (HS), and silencing of OGT expression attenuates this effect. Lanes 1, 2, and 3 are control cells transfected with either no siRNA, noncoding (NC) siRNA, or OGT siRNA, respectively. OGT siRNA decreases basal levels of O-glycosylated intracellular proteins. Lanes 4, 5, and 6 are non-HS GLN-treated cells (transfected similarly). Cells showed increased O-glycosylation levels with GLN treatment even in the absence of stress. OGT silencing attenuates this GLN mediated O-glycosylation (lane 6). Lanes 7, 8, and 9 are HS control cells (HS CT) and lanes 10, 11, and 12 are HS with GLN (with the same transfections). HS increases O-glycosylation, and GLN treatment again enhances this effect. OGT silencing attenuates GLN-induced O-glycosylation, whereas NC oligos do not. Western blot is representative of 3 separate experiments. MW, molecular weight.