(A, B) Depletion of endogenous FBXO11 in carcinoma cells stabilizes Snai1 protein and causes gene expression changes characteristic of EMT. MCF7 and HCT116 cells were transduced with lentiviral shRNAs targeting FBXO11 or empty vector pLKO. Cells were collected and subjected to immunoblotting and RT-qPCR analysis for indicated epithelial and mesenchymal markers.
(C) Depletion of FBXO11 induces EMT. Images of control and FBXO11-depleted HCT116 cells show morphological differences.
(D) Depletion of FBXO11 enhances cell migration. Representative images of wound (scratch)-healing assay demonstrate FBXO11-depleted HCT116 cells migrated faster than control cells, as shown 48 hours after scratching.
(E) Depletion of FBXO11 stimulates cell invasiveness in vitro. In the Transwell invasion assay, few control HCT116 cells invade and migrate through the matrix layer. By contrast, FBXO11-depleted HCT116 exhibit dramatically improved invasive potential.
(F) Depletion of FBXO11 enhances tumor invasion in transplant assay. Control and FBXO11-depleted HCT116 cells were injected into immunodeficient mice. Tumors were isolated and sectioned for H&E staining. Dashed line denotes the tumor boundary.