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. 2011 Aug 12;286(39):34286–34297. doi: 10.1074/jbc.M111.259457

FIGURE 6.

FIGURE 6.

PUGNAc, an inhibitor of O-GlcNAcase, increases the binding of 4E-BP1 to eIF4E in perfused liver. Livers from non-diabetic Ins2+/+ mice were perfused in situ with a nonrecirculating medium containing either 40 μm PUGNAc (PUG) or vehicle alone. A, O-GlcNAcylation of total 4E-BP1 was assessed by treating supernatants from liver WCL with λ-phosphatase (PPase) followed by Western blot analysis. O-GlcNAcylation of full-length 4E-BP1 (B) and truncated 4E-BP1 (tr4E-BP1; C) bound to eIF4E was measured by Western blot analysis. D, the association of 4E-BP1 with eIF4E was examined by immunoprecipitating eIF4E from liver supernatants and measuring the amount of 4E-BP1 in the immunoprecipitate by Western blot analysis. E, total eIF4E, 4E-BP1, and 4E-BP1 O-GlcNAcylation was assessed in WCL by Western blot analysis. Representative blots are shown. Values are means + S.E. (n = 8). Statistical significance is denoted by *, p < 0.05.