FIG 3.
The 4EBP1-eIF4E complex interacts with GCNT3 to maintain GCNT3 stability. (A) eIF4E protein levels were inhibited in EBVaGC cells compared with EBVnGC cells. (B) Western blot analysis of eIF4E in SGC7901-LMP2A cells and in GT38 cells transfected with LMP2A-siRNA for 48 h. (C) GCNT3 protein levels after knockdown of eIF4E for 48 h (50 nM) in BGC823 and SGC7901 cells. (D) Coimmunoprecipitation in SGC7901 cells followed by Western blotting showed that GCNT3 interacted with eIF4E and 4E-BP1. (E) Coimmunoprecipitation and Western blot analysis showed that LMP2A inhibited the interaction between GCNT3 and eIF4E in SGC7901 cells. (F) Comparison of GCNT3 degradation in SGC7901 cells infected with siNC or sieIF4E and then treated with control (DMSO) or 10 μg/ml CHX in a time series. (G) Analysis of panel F in triplicate. (H) Comparison of GCNT3 degradation transfected with siNC or sieIF4E in SGC7901 cells and without or with treatment with MG132 (20 μM). All experiments were performed thrice. Error bars indicate SD (n ≥ 3). *, P < 0.05; **, P < 0.01.