Co-immunoprecipitation of Protein Complexes from Different Subcellular Compartments in Vasculogenic Mimicry Studies.

ABSTRACT

Aberrant extravascular expression of VE-cadherin has been observed in metastasis associated with vasculogenic mimicry (VM); we have recently shown that in VM prone cells VE-cadherin (mainly in the form of phospho-VE-cadherin in Y658) is in part located in the cell nucleus, which associates with p120-catenin and the transcription factor kaiso allowing increased plasticity that potentiates VM development in malignant cells. In this chapter, we describe the protocol to analyze protein-protein interactions in subcellular fractions with particular focus in VE-cadherin. The verification of the subcellular interactome of VE-cadherin and other key proteins involved in VM shed light to novel functions of endothelial proteins aberrantly expressed in tumor cells and their consequences in cell plasticity during VM development.