Fig. 1. The mTORC1 substrates 4EBP1/2 prevent cell death in response to glucose starvation in human, mouse and yeast cells.

A, B The indicated cell lines were grown in glucose starved medium (Glc strv) and treated with vehicle (V), Rapamycin (Rap), KU-0063794 (KU), cycloheximide (CHX) or 4EGI for 48 h. Cell death was measured by PI staining and flow cytometry. C Scheme of the mTORC1 downstream signaling pathways controlling mRNA translation initiation and the impact of the inhibitors used in (A, B). D The indicated cell lines were grown in complete media or starved for glucose (Glc) for 48 h. Cell death was analyzed as in (A, B). The level of the indicated proteins was analyzed by immunoblotting. E The indicated cell lines were grown in glucose starved medium (Glc strv) for 48 h. Cell death and protein levels were analyzed as in (A, B) and (D) respectively. F The indicated cell lines were grown in complete media or starved for glucose (Glc) for 48 h. Cell death and protein levels were analyzed as in (A, B) and (C) respectively. G The indicated cell lines were glucose (Glc) starved for 48 h. Med8a and HD-MB03 cells were treated with 1 μg/ml doxycycline for 72 h. Cell death was measured as in (A, B). H 4E KO MEF were transfected with control siRNA (scr) or siRNAs targeting Eif4e and grown in glucose starved medium (Glc strv) for 48 h. Cell death and protein levels were analyzed as in (A, B) and (D) respectively. I WT, caf20Δ, eap1Δ, or eap1Δ/caf20Δ S. cerevisiae strains were plated by serial dilution on solid complex medium with (YPD) or without (YP) 2% glucose at 37 °C. J WT or eap1Δ strains were grown in liquid medium containing no glucose (YP) for 2 weeks at 30 °C and were plated by serial dilutions onto complete YPD agar plates. Data are shown as the mean ± SD. Statistics: unpaired one-sided Student’s t test (A, B, D, E, F, G, H); n = 3 independent experiments for (A, B, D, E, F, G, H). Source data are provided as a Source Data file.