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. Author manuscript; available in PMC: 2020 Sep 19.
Published in final edited form as: Mol Cell. 2019 Sep 4;75(6):1218–1228.e6. doi: 10.1016/j.molcel.2019.07.027

Figure 2. Translation initiation status during 2–5AMD.

Figure 2.

(A) Activity of mTOR kinase monitored by western blot for phosphorylated 4E-BP1. Matching translational activity was measured by 35S metabolic labeling. The small molecule mTOR inhibitor INK128 is used as a control. The 2–5A and INK128 treatments were for 2 h. (B) Quantification of the gels in (A). Error bas are S.E. from the two series in (A). (C) Western blot analysis of core components of the cap-binding eIF4F complex: eIF4E, eIF4A and eIF4G. Data from control (Flag) and eIF4E immunoprecipitation (IP) experiments are shown with and without 2–5A treatment. (D) BioAnalyzer NanoChip profiling of rRNA in the samples in (C). (E) Mass spectrometry analysis of proteins that co-immunoprecipitate with the cap-binding translation initiation factor eIF4E (Figure S2, Table S1).