4egi 1

JNK inhibitor SP600125 (SP), eIF2α phosphatase enzymes inhibitor salubrinal (Sal) and mTOR inhibitor rapamycin (Rap) were purchased from Tocris Bioscience (Bristol, UK). p70S6K inhibitor PF-4708671 (PF) was purchased from Selleck Chemicals (Houston, TX, USA). AP-1 inhibitor curcumin, cell counting kit-8 (CCK8) and ER stress inducer tunicamycin (Tun) were purchased from Sigma (Lyon, France). The eIF4E/eIF4G interaction inhibitor 4EGI-1, mTOR siRNA, GFP siRNA, JNK siRNA, GRP78 siRNA, ATF4 siRNA and antibodies against GRP78, eIF2α and β-actin were purchased from Santa Cruz Biotechnology (Heidelberg, Germany). Antibodies against phospho-eIF2α (Ser-51), phospho-p70S6K (Thr-389), phospho-mTOR (Ser-2448), phospho-Raptor (Ser-863), phospho-c-Jun (Ser-73), phospho-JNK (Thr-183/Tyr-185), phospho-4E-BP1 (Thr-37/46), p70S6K, mTOR, Raptor, c-Jun, JNK, 4E-BP1 and ATF4 were purchased from Cell Signaling Technology (Danvers, MA, USA).

The following 43 reagents were used in this study: AMG 102 and panitumumab (Amgen); cercosporamide (BioAustralis); AKT inhibitor V (Calbiochem); AS-252424, bisindolylmaleimide, CGP 57380 and imatinib (Cayman Chemical); CTX ILK inhibitor (CRC); Avastin (bevacizumab) (Genentech/Roche); 4EGI-1, ABT-737, ABT-737 enantiomer, pazopanib, and retinoic acid (Santa Cruz Biotechnology); erlotinib, lapatinib, sorafenib, and temsirolimus (Scientifix); bortezomib and CYT387 (Selleck Chemicals); AKT-I-1/2, axitinib, bicalutamide, cilostazol, cyclopamine, DAPT, dasatinib, docetaxel, doxorubicin, floxuridine, fluorouracil, goserelin, ifosfamide, PD98059, ribavirin, salinomycin, SU11274, sunitinib, Tamoxifen, and vinblastine (Sigma); NSC 7908 (Tocris); and temozolomide (Wyeth).

HeLa and HEK293 cells were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (Life Technologies, Carlsbad, CA, USA) according to ATCC guidelines. Cells were treated with proteasome inhibitor MG132 (Merck, Kenilworth, NJ, USA) and 4EGi-1 (Santa Cruz Biotechnology, TX, USA), which inhibits the interaction of the translation initiation factors eIF4E and eIF4G, to specifically block eIF4E-dependent translation^{17 (link)}. To confirm the stability of mRNAs expressed from β-globin constructs, cells were treated with Actinomycin D (ActD; Sigma, St. Louis, MO, USA) to inhibit transcription.