Fig. 4

CHIP triggers the EMT of DLD-1 cells via inactivating GSK-3β. a The mRNA expression of CDH1, EpCAM, CK8, and CK18 in the established DLD-1 transfected cell lines, including siCHIP and sictrl, as well as hCHIP and ctrl cells. β-actin normalized gene expression, measured in triplicates was displayed. Significant differences were indicated (Student’s t test, **P < 0.01, ***P < 0.001). b The protein expression of E-cadherin, EpCAM, and CK8/18 in established DLD-1 transfected cell lines were analyzed by western blotting. The level of each protein was normalized against Actin. c Representative IHC images for CHIP, E-cadherin, and EpCAM expression in the established DLD-1 transfected cell lines (×200 magnification). d Western blotting analysis of the protein expression of GSK-3β, p-GSK-3β, Slug, and E-cadherin in the DLD-1 established transfected cell lines. The level of each protein was normalized against Actin. e–g Hierarchical clustering analysis on the differentially expressed proteins among DLD-1 CHIP-overexpressing and control cells, which related to the CRC disease and cell–cell adhesion and junction based on the disease, GO, and KEGG pathway enrichment analysis, respectively. CDH1 is marked by a red box