Ribo-seq of Eif6 WT and Eif6 S235A Mouse Embryonic Fibroblasts (MEFs) pre-treated with harringtonine
Ribo-seq of Eif6 WT and Eif6 S235A Mouse Embryonic Fibroblasts (MEFs) pre-treated with harringtonine
Immortalized MEF cells were pre-treated with 2ug/ml harringtonine for 2 minutes followed by 100ug/ml cyclohexamide for other 2 minutes. Finally, the cells were washed once with ice-cold PBS 10ug/ml cyclohexamide. Cells were lysed in 20mM Tris-HCl, 50mM NaCl, 10mM MgCl2, 1mM DTT, 100ug/ml cyclohexamide, 1% NP-40, 0.2mg/ml heparin. Ribosome Protected Fragments (RPF) were obtained following Ingolia 2017 (10.1016/j.ymeth.2017.05.028).  Immagina Biotechnology LACEseq #LS-001_12