Western Blotting Analysis of Cell Signaling Pathways

Western blotting analysis was performed using the same procedure demonstrated previously^{7 (link)}. Briefly, the radioimmunoprecipitation assay lysis buffer (Beyotime, China) was used to extract total cell protein, which was then quantified using the bicinchoninic acid protein assay kit (Beyotime, China) and resolved on a 6–12% sodium dodecyl sulphate-polyacrylamide gel, and then incubated with relevant primary antibodies. GAPDH and histone (ZSGBBIO, China) were used as loading controls. The following primary antibodies were utilised in this research: anti-PPARδ (Proteintech), anti-CXCR4 (Proteintech, China), anti-vimentin (Proteintech), anti-GAPDH (Proteintech), anti-β-catenin (Proteintech), anti-Slug (Proteintech) and anti-N-cadherin (Proteintech). We used GSK3787 (Abcam) and XAV-939 (Abcam, UK) as PPARδ and β-catenin inhibitors, respectively. The enhanced chemiluminescence system (Bio-Rad, Hercules, EDA USA) was used to detect protein expression levels and Scion imaging software was used to capture images.