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. 2012 Apr 4;287(22):18500–18509. doi: 10.1074/jbc.M111.327528

FIGURE 4.

FIGURE 4.

Down-regulation of eIF3g inhibits NMD. HeLa cells were depleted of eIF3b, eIF3c, eIF3g, or Upf1 using specific siRNAs. Two days later, cells were retransfected with (i) pmCMV-Gl either Norm or Ter, (ii) pmCMV-GPx1 Norm or Ter, (iii) phCMV-MUP, and (iv) pRL-CMV. A, Western blotting demonstrating the efficient down-regulation of each indicated protein. B, RT-PCR of Gl mRNAs. The levels of Gl mRNAs were normalized to the levels of MUP mRNAs. The normalized levels of Gl Norm mRNA in the presence of each siRNA were set to 100%. C, RT-PCR of GPx1 mRNAs. As in B, except that the GPx1 mRNAs were analyzed by RT-PCR. D, RT-PCR of RLuc mRNAs. The levels of RLuc mRNAs were normalized to the levels of endogenous SMG7 mRNAs. The normalized level of RLuc mRNA in the cells co-transfected with pmCMV-Gl Norm, pmCMV-GPx1 Norm, and Control siRNA was set to 100%. E, translational efficiency of RLuc mRNAs. RLuc activity was normalized to the level of RLuc mRNA. Translational efficiency of RLuc mRNA in the cells co-transfected with pmCMV-Gl Norm, pmCMV-GPx1 Norm, and Control siRNA was set to 100%.