GSM9068647: sh-NC_2-IP; Homo sapiens; RIP-Seq
acRIP assay was conducted using the GenSeq ac4C RIP kit (GS-ET-005, Cloudseq Biotech, Shanghai, China). Total RNA was extracted and fragmented into 100–200 bp segments, with a portion reserved as an input control. Magnetic beads conjugated with ac4C antibodies were used to immunoprecipitate ac4C-modified RNA from the fragmented samples. The purified RNA was subsequently analyzed by RT-qPCR to measure CHAF1A expression levels, normalized to the input control. For acRIP sequencing, the purified RNA was utilized to construct RNA libraries using the GenSeq® Low Input Whole RNA Library Prep Kit (GenSeq, Inc.), and quality control was performed with an Agilent 2100 Bioanalyzer. High-throughput sequencing was then conducted on an Illumina NovaSeq 6000 platform.