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. 2014 Sep 2;5:4790. doi: 10.1038/ncomms5790

Figure 7. Role of non-canonical motifs in translational repression.

Figure 7

(a,b) S2 cells were transfected with plasmids expressing a F-Luc reporter (F-Luc) and HA-CUP (N-Term, residues 1–402) or HA-Thor (full length, either WT or the indicated mutants). The F-Luc mRNA levels were analyzed by northern blotting and were normalized to those of the control RNA. The normalized F-Luc mRNA levels were used to normalize the F-Luc activity to obtain the translation efficiencies, which were set to 100 in cells expressing the HA peptide. The mean values±s.d. from three independent experiments are shown. The red, dashed lines indicate the F-Luc levels for the maximum repressive activity that was exhibited by the 4E-BPs. The black, dashed lines indicate the F-Luc levels expected in the absence of repression. (c,d) Upper panels show northern blot (NB) analyses of representative RNA samples corresponding to the experiments that are shown in a,b, respectively. The lower panels show the expression of the 4E-BP proteins analyzed by WB. The original northern and WB can be found in Supplementary Fig. 9.