Detection of N4-Acetylcytidine(ac4C) in human mRNA using RedaC:T-Seq in SK-HEP-1 Cells. Homo sapiens

In this study, we employed RedaC:T-seq, a chemical conversion and high-throughput sequencing approach, to systematically detect ac4C modifications in human SK-HEP-1 cells. To assess the role of NAT10 in ac4C deposition, we generated NAT10-knockdown SK-Hep-1 cell lines using short hairpin RNA (shRNA) technology, with shNC (non-targeting control) serving as a negative control. The efficiency of NAT10 knockdown was confirmed through quantitative PCR (qPCR) and Western blot (WB) analyses. Our results confirm the presence of ac4C and reveal a marked reduction of ac4C levels following NAT10 knockdown.