5 A–H). Then, qRT-PCR (Figure. 5I–L) showed DNMT3B and NSUN6 were significant DEGs between HF and control rat samples (n = 6), consistent with bioinformatics results, while DNMT1 and NOP2 did not. The results of western blot illustrated the significance of NSUN6 (Figure. 5 M). Altogether, the four-gene m5C-based model exhibited excellent diagnostic capability for HF. Additionally, the ssGSEA analysis of NSUN6 showed Hedgehog signaling pathway was enrichment in HF samples, suggesting its potential role in the progression of HF (Figure. 5 N-O). Here, DNMT1 is linked to hydrops fetalis.