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. 2025 Jul 28;21(11):4997–5014. doi: 10.7150/ijbs.113325

Figure 3.

Figure 3

NAT10 acetylates lncRNA XIST to promote VEGFA expression. (A) Volcano plots of differentially acetylated genes identified by acRIP-seq. (B) The level of VEGFA protein in CM collected from shXIST GC cells was measured by ELISA. (C) Distribution of NAT10-binding regions and ac4C peaks on XIST lncRNA visualized by IGV. (D) The acRIP-qPCR assays were performed to detect the ac4C level of XIST in GC cells treated with shNAT10 or NAT10 plasmid. (E) The RIP assays were performed to detect the interaction between NAT10 and XIST in AGS and MGC-803 cells. (F) The interaction between NAT10 and XIST was analyzed by RIP-qPCR in GC cells with NAT10 knockdown or overexpression. (G) The expression level of XIST was detected by qPCR in GC cells treated with shNAT10s or NAT10 plasmid. (H) The mRNA stability was detected by qPCR in AGS and MGC-803 cells treated with actinomycin D (5 μg/ml). (I) Schematic representation of NAT10-targeted ac4C motifs within XIST (upper panel). The ac4C sites within the exon of XIST were mutated and inserted into the pmirGLO plasmid for dual-luciferase assays (lower panel). (J) Dual-luciferase assays were performed in shNAT10 or NAT10-overexpression GC cells using pmirGLO-XIST WT or pmirGLO-XIST mut plasmid.