GSM8409054: Pnx-sham, Input; Mus musculus; RIP-Seq
For mouse m6A RNA-IP-seq, we started with ~200–300μg of total RNA of CD31+CD45- ECs isolated from lung 0 day and 7 days after PNX, using magnetic beads. The NEBNext® Ultra II Directional RNA Library Prep kit was used to construct RNA sequencing libraries for Input RNA samples without immunoprecipitation, as well as IP RNA samples after immunoprecipitation. m6A RNA-Seq service was provided by Cloudseq Biotech Inc. (Shanghai, China). Briefly, m6A RNA immunoprecipitation was performed with the GenSeqTM m6A RNA IP Kit (GenSeq Inc., China) by following the manufacturer's instructions. 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. Briefly, Paired-end reads were harvested from Illumina HiSeq 4000 sequencer, and were quality controlled by Q30.