The m6A demethylase FTO promotes C/EBPβ-LIP translation to perform oncogenic functions in breast cancer cells (RNA-sequencing)
The m6A demethylase FTO promotes C/EBPβ-LIP translation to perform oncogenic functions in breast cancer cells (RNA-sequencing)
Cells were grown as usual and harvested at 80% confluency in ice cold PBS with 100 μg/ml cycloheximide. RNA was isolated from a portion of cells processed for ribosome profiling using the RNeasy kit (Qiagen).  mRNA was isolated by NEXTFLEX® Poly(A) Beads (PerkinElmer) and quality checked by RNA 6000 nano chip (Agilent) on a Bioanalyzer 2100 (Agilent). Sequencing libraries were prepared using NEXTFLEX® Rapid Directional qRNA-Seq™ Library Prep Kit (PerkinElmer) according to the manufacturer's instructions. Library quality was confirmed using a Bioanalyzer 2100 (Agilent) with the DNA 7500 kit (Agilent) and libraries were quantified by Qubit dsDNA BR assay (ThermoFisher).