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

FIGURE 2.

FIGURE 2.

CTIF is an essential protein for recruiting eIF3 into CT complex. A, IPs of FLAG-CTIF. HeLa cells were transiently transfected with plasmid, either pcDNA3-FLAG or pcDNA3-FLAG-CTIF. Cell extracts were then either untreated (−) or treated (+) with RNase A and subjected to IPs using α-FLAG-conjugated resin. Western blotting was carried out to detect the indicated proteins (upper panel). Cellular protein β-actin served as a negative control. The complete digestions of endogenous RNAs by RNase A treatment were demonstrated by RT-PCR (lower panel). B, IP of FLAG-eIF3g. As in A, except that cells were transfected with either pcDNA3-FLAG or pcDNA3-eIF3g. C, IP of Myc-CBP80. The extracts of HeLa cells that were either undepleted (Control siRNA) or depleted of endogenous CTIF (CTIF siRNA) and were transiently transfected with either pCMV-Myc or pCMV-Myc-CBP80 were subjected to IP using α-Myc antibody. D, IP of Myc-eIF4E. As in C, except that cells were transfected with either pCMV-Myc or pCMV-Myc-eIF4E.