Fig. 5: CPNE7 interacts with NONO to promote ZFP42 transcription.

A Volcano plots of DEGs in shControl group vs shCPNE7 groups. B, C GO and KEGG analysis of DEGs from left volcano plot (B) and right volcano plot (C) in (A). D GSEA analysis of RNA-sequencing data from shControl group and shCPNE7-1 group. E Venn plot of HumanTFDB and DEGs in RNA-sequencing (|FoldChange | > 1.5). F 5 genes exhibiting the same trend in RNA-sequencing and qRT-PCR. G Immunofluorescence staining of CPNE7 and NONO in HCT116 cells. Scale bar: 10 μm. H Dual-luciferase reporter assay were performed in shControl and shCPNE7 293 T cells. I Proteins from the IP assay on SW480 cells were separated by SDS-PAGE and detected by silver staining. J, K Interaction between NONO and CPNE7 were verified in 293 T (J) and HCT116 (K) cells by Co-IP assay. L The effect of CPNE7 and NONO on ZFP42 promoter activity was detected by dual-luciferase reporter assay. M Left panel: schematic diagram of the structure of truncated ZFP42 promoter reporter plasmids. Right panel: Changes in luciferase activity following truncation at different positions of the ZFP42 promoter. N ZFP42 mRNA expression were detected in shControl and shNONO SW620 cells. O Expression of NONO and ZFP42 were detected in CPNE7 knockout HCT116 cells by Western blot. P, Q mRNA expression level (P) and protein expression level (Q) of ZFP42 in shControl and shZFP42 HCT116 cells. R Cell viability was measured by CCK-8 assay. S Colony formation assays of shControl and shZFP42 HCT116 cells. Representative images are shown on the left, and the statistical analysis is shown on the right. T Transwell assays of shControl and shZFP42 HCT116 cells. Representative images are shown on the left, and the statistical analysis is shown on the right. U Wound-healing assays of shControl and shZFP42 HCT116 cells. Representative images are shown on the left, and the statistical analysis for migration rates is shown on the right. V Apoptosis detection assays of shControl and shZFP42 HCT116 cells. Representative images are shown on the left, and the statistical analysis for apoptotic rates (including early apoptosis and late apoptosis) is shown on the right. For (F–V), data are shown as mean ± SD and two-tailed unpaired Student’s t-test was used. *p < 0.05, **p < 0.01, ***p < 0.001. Data are representative of at least three independent experiments.