Figure 2. Identification of non-canonical motifs in Thor and 4E-T.

(a) WB showing the interaction of HA-Thor (full length, either WT or mutated) with endogenous eIF4E in S2 cells. The proteins were immunoprecipitated using anti-HA antibodies. The inputs (1%) and immunoprecipitates (30%) were analyzed by WB using anti-HA and anti-eIF4E antibodies. The original WB shown in this panel can be found in Supplementary Fig. 8. (b) WB showing the interaction of GFP–4E-T (full length, WT or mutated) with endogenous eIF4E. The proteins were immunoprecipitated using anti-GFP antibodies. The inputs (1%) and immunoprecipitates (30%) were analyzed by WB using anti-GFP and anti-eIF4E antibodies. The original WBs can be found in Supplementary Fig. 8. (c) MBP pull down showing the interaction of His₆-tagged eIF4E (residues 69–248, trunc) with MBP-tagged 4E-BP C+NC fragments (see Supplementary Table 1) and eIF4G (residues 578–650). All the fragments contained, in addition, a C-terminal GB1 tag. The input samples (10%) and bound fractions (15%) were analyzed using SDS–PAGE followed by Coomassie blue staining. The size markers (kDa) are shown to the right of each panel.