biostudies-arrayexpressMus musculushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-13335Analysis of global variability of translation initiation between the Eif6 WT control genotype and the Eif6 S235A mutated MEF lines pre-treated with harringtonine. Ribosome Protected Fragments (RPFs) were obtained following the method reported in Ingolia NT et al., NatProtoc 2012. (doi:10.1038/nprot.2012.086)biostudies-arrayexpressLibrary Construction - Immagina Biotechnology LACEseq #LS-001_12Nucleic Acid Extraction - 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).Sample Collection - 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.Sequencing - 500 pM library, Phix 10%, NextSeq 1000/2000 P2 100 cycles Illumina Kit, sequenced with NextSeq 2000MINSEQE ScoreAssays and DataProcessed DataorganisationMAGE-TAB FilesData Transformation - The raw fastq files underwent trimming and UMI extraction as reported in the Laceseq kit guidelines. Reads aligning to tRNAs and ncRNAs were removed using bowtie2MetabolomicsUnknownTranscriptomicsGenomicsProteomicsNextSeq 2000Ribo-seqMus musculusERP151032Ivan FerrarifalseRibo-seq of Eif6 WT and Eif6 S235A Mouse Embryonic Fibroblasts (MEFs) pre-treated with harringtonineAnalysis of global variability of translation initiation between the Eif6 WT control genotype and the Eif6 S235A mutated MEF lines pre-treated with harringtonine. Ribosome Protected Fragments (RPFs) were obtained following the method reported in Ingolia NT et al., NatProtoc 2012. (doi:10.1038/nprot.2012.086)2026-02-10T00:00:00Z2026-05-27T12:52:57.877Z2023-09-11T15:13:32.158ZE-MTAB-13335ERP151032EFO_0002944EFO_0004170EFO_0008891EFO_0005518EFO_0003816EFO_0004184