Fig. 3
Screening of genes with abnormal m5C hypermethylation. (A) m5C dot blot assays using mRNA from MCF7 and T47D cells with or without NSUN2 knockdown. Methylene blue staining was used as a loading control. (B) Volcano plot showing differentially expressed m5C sites between tumor and adjacent tissues (n = 5). (C) Different proportions of m5C modifications in mRNA regions between BC tissues and adjacent tissues. (D) KEGG analysis revealed that m5C-hypermethylated genes with high expression levels in BC tissues were enriched in oncogenic signaling pathways. (E) Overlap of target genes from RNA-Seq and RNA-BisSeq of BC tissues (n = 5). The horizontal dashed line represents the threshold of the adjusted p-value of 0.05, while the vertical line represents the threshold of a log2-fold change greater than 1.5. (F) Clustering analysis heatmap showing the display level of different tissue samples. (G) Database analysis revealed an increase in HGH1 expression in tumor tissue. (H) Survival analysis showed that the group with high expression of HGH1 had a worse prognosis. (I) and (J) RT-qPCR and IHC experiments proved that HGH1 was expressed at higher levels in BC tissues than in paired adjacent tissues. Data are mean ± SD of three independent experiments for (I). *p ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001 by t test or by log-rank (Mantel-Cox) test