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. 2023 Mar 15;114(6):2596–2608. doi: 10.1111/cas.15764

FIGURE 6.

FIGURE 6

Identification of ELF3‐regulated gene signature and clinical relevance. A, Immunoblotting for ELF3 in ECC4, A99, and DMS53 cells treated with the negative control (siNC) or siELF3. GAPDH was used as a loading control. B, Flowchart for extracting ELF3‐regulated genes by combining cleavage under targets and tagmentation (CUT&Tag) for ELF3 and ELF3 knockdown RNA‐seq. C, Bar chart showing the combined scores calculated by EnrichR pathway analysis (MSIgDB Hallmark) of 232 ELF3‐regulated genes. D, IGV visualization of ELF3 CUT&Tag signals for representative five ELF3‐regulated genes (AURKA, CDC25B, CLDN4, ITGB6, and YWAHB). E, Bar chart showing cell viability at 96 h after ELF3 knockdown by siRNA in ECC4, A99, and DMS53 cells. Data are representative of three independent experiments. **p < 0.01. F, Bar chart showing colony formation in soft agar at day 7 in the ELF3 knockout clone of ECC4 cells. Data are representative of three independent experiments. *p < 0.05. G, Heatmap showing ELF3‐regulated 232 genes in 34 neuroendocrine carcinomas of the gastrointestinal system (GIS‐NECs). Unsupervised clustering identified two clusters: ELF3‐sig–high GIS‐NEC and ELF3‐sig–low GIS‐NEC. H, Enrichment plot of GSEA with 34 GIS‐NEC gene expression data