EpEX-specific (rat anti-mouse EpCAM G8.8, BD Biosciences, Heidelberg, Germany) and vimentin-specific antibodies (rabbit monoclonal to vimentin EPR3776, Abcam, Cambridge, UK) were used for IHC. Immunostaining was performed using the avidin-biotin-peroxidase method (VECTASTAIN, Vector Laboratories, Burlingame, CA, USA) according to the manufacturers’ protocol. IHC intensity scores (IHC score) were calculated as the product of intensity (0 to 3+) and the percentage of expressing tumor cells (score: 0 = 0 to 5%, 1 = 5 to 25%, 2 = 25 to 50%, 3 = 50 to 75%, and 4 = 75 to 100%). IHC scores represent averages of values independently assessed by a minimum of two experimenters, who were blinded to sample identity.

EMT scores were calculated as the percentage of mesenchymal, spindle-like cells (0 to 100%) and the level of cell-cell contact (disseminated cells represent 1 = 0 to 25%, 2 = 25 to 50%, 3 = 50 to 75%, and 4 = 75 to 100%). Three experimenters scored independently each cell line at an average confluence of 50 to 80% for three independent passages.

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