Fig. 4: ATXN1 facilitate EMT by acting as a TF of CDH2 (N-cadherin). | Oncogene

Fig. 4: ATXN1 facilitate EMT by acting as a TF of CDH2 (N-cadherin).

From: Targeted methylation of cg24067911 suppresses colorectal cancer metastasis through BCL6-ATXN1-CDH1 axis

Fig. 4

A GO/KEGG analysis of cg24067911 methylation status and ATXN1 expression level. Left, box plot of common GO terms enriched by DEGs of cg24067911 hypermethylation (dCas9-DNMT3a treated DLD-1 cells versus controlled DLD-1 cells); Right, box plot of common GO terms enriched by DEGs of ATXN1 (low versus high expression). GO, Gene Ontology. B GSEA plot of ATXN1 functioning enriched in Hallmark EMT pathway. C Immunoblotting by western blot of ATXN1 and EMT markers E-cadherin and N-cadherin in ATXN1-knockdown HCT116 by siRNA-ATXN1 and ATXN1-overexpressing RKO. D, E RT-qPCR data of ATXN1, CDH1 and CDH2 in ATXN1-knockdown HCT116 D and ATXN1-overexpressing RKO E Student’s t-test. F Protein-protein interaction analysis of ATXN1 and EMT-associated genes. G The binding ability of ATXN1 to CDH2 promoter sequence. Left up, the plot of DNA agarose gel electrophoresis of the ChIP assay. Left down, the predicted binding motif. Right, quantified data of PCR of the immunoprecipitated DNA from ChIP assay. H The dual luciferase reporter assay measured transcriptional regulatory activity of ATXN1 on CDH2 promoter with wild type or mutations. Left, the schematic plot of CDH2 promoter and designed mutations in promoter plasmids. Right, measurement and comparisons of relative luciferase activity of CDH2 promoter in wild type or mutated type between ATXN1 overexpression and normal control. Student’s t-test. The data are shown as the mean ± SD; *P < 0.05, **P < 0.01, and ***P < 0.001.

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