mTEC cells transfected with si-circ-RNF216 or siRNA-NC

The mTEC cells underwent transfection with siRNA-2 or siRNA-NC. RNA was first enriched using magnetic beads conjugated with Oligo (dT), after which the mRNA was randomly fragmented by the addition of Fragmentation Buffer. Taking the fragmented mRNA as the template, the first-strand and second-strand cDNA were synthesized sequentially, followed by cDNA purification. The purified double-stranded cDNA then underwent end repair, A-tailing, and sequencing adapter ligation; subsequently, fragment size selection was performed using AMPure XP beads. Finally, the cDNA library was obtained through PCR enrichment. The generated libraries were examined by Qsep400 and subjected to sequencing on an Illumina HiSeq2000 instrument. Next, the clean data was obtained after raw data filtering, and aligned to the mm10 mouse reference genome.