Epitranscriptomics m6A analyses reveal distinct m6A marks in tumor necrosis a (TNF-a)-treated HeLa cells

We have performed a epitranscriptomics study in which we first tretaed TNF-alpha in HeLa cells and DMSO was used as a negative control. Total RNA was isolated from TNF-alpha treated cells as well as control cells and apoptotic rate was determined by flow cytometry. Total RNAs were subjected to miCLIP to identify differentially m6A methylated RNAs. We then transcriptionally analyzed apoptotic genes which have differential m6A methylation in TNF-alpha tretament by qPCR. Afterthat, candidate genes were examined at the level of translation by polysome fractioantion assay. Overall design: Total RNA was isolated from negative control (DMSO) and TNF-alpha treated HeLa cells in triplicate to perform miCLIP assay.