5-methylcytosine modification profiling of messenger RNA in breast cancer brain metastasis
We performed m5C MeRIP-seq to clarify the role of m5C RNA modification in the pathogenesis of BCBM Overall design: Sample: cell line; Group: breast cancer brain metastasis (BCBM) vs breast cancer (BC),each with 3 replicates; Methods:m5C RNA-Seq service was provided by Cloudseq Biotech Inc. (Shanghai, China). Briefly, m5C RNA immunoprecipitation was performed with the GenSeqTM m5C RNA IP Kit (GenSeq Inc., China) by following the manufacturer's instructions. Both the input sample without immunoprecipitation and the m5C IP samples were used for RNA-seq library generation with NEBNext® Ultra II Directional RNA Library Prep Kit (New England Biolabs, Inc., USA). The library quality was evaluated with BioAnalyzer 2100 system (Agilent Technologies, Inc., USA). Library sequencing was performed on an illumina Hiseq instrument with 150bp paired-end reads.