Fig. 6

SLCO4A1-AS1 activates c-Myc signalling in a Cdk2-dependent manner in CRC. a Gene expression profiles of CRC cells transfected with SLCO4A1-AS1 plasmids (oeSLC) or siSLCO4A1-AS1 (siSLC). b GSEA focused on a set of signalling pathways after SLCO4A1-AS1 overexpression and SLCO4A1-AS1 silencing, summarized based on the enrichment score. c Functional annotation clustering of common DEGs regulated by SLCO4A1-AS1 in two independent RNA sequencing assays is shown. The x-axis shows the ratio of the number of genes in a given category of functional annotations divided by the total number of DEGs. The y axis shows categories of functional annotations. d GSEA showed that high SLCO4A1-AS1 expression was positively related to the activation of c-Myc signalling. e Expression heatmap of the target genes of c-Myc signalling shown in two independent RNA sequencing assays. f Western blotting showing that shRNA-induced SLCO4A1-AS1 knockdown decreased the protein levels of Cdk2 in SW620 and HT29 cells. Histone (Histone H3.1), nuclear marker; GAPDH, cytoplasmic marker; Nuc, Nucleus; Cyto, Cytoplasm. g The c-Myc proteins were analyzed by western blotting using phospho (Ser62)- and pan-Myc antibodies in CRC cells transfected with control or SLCO4A1-AS1 (shSLC or oeSLC) plasmids. h Western blotting of proteins from the control or shSLCO4A1-AS1 group was performed in HCT116 and LoVo cells with or without CVT-313 treatment. i The mRNA levels of downstream target genes of the c-Myc pathway were measured by qRT-PCR in CRC cells transfected with control and SLCO4A1-AS1-depleted plasmids or cotransfected with Cdk2 overexpression plasmids. j Luciferase assays were performed to assess the effects of SLCO4A1-AS1 and Cdk2 on the c-Myc pathway. k Cell viability was measured by CCK-8 assays in CRC cells. l FACS analyses of the effects of SLCO4A1-AS1 on the cell cycle in HCT116 and LoVo cells. *P < 0.05; **P < 0.01; ***P < 0.001