Figure 3

ALK Gene Expression of Pre-Clinical Models of IBC. A. Analysis of gene expression levels of ER/PR/Her2, EGFR and ALK in each of the 7 available pre-clinical models of IBC including SUM149, Mary-X, FC-IBC01, FC-IBC02, SUM190, MDA-IBC-3 and KPL-4 cells and non-IBC human breast tumor cell lines MDA-MB-231, SUM159 and MCF-7. The highest levels of ALK gene expression was detected in the triple negative IBC cell lines FC-IBC01, FC-IBC02 and Mary-X, which each recapitulate the formation of IBC tumor emboli in vivo. B. Light micrograph of histology of FC-IBC01 xenograft showing poorly differentiated tumor with high nuclear grade and prominent mitotic activity, with visible invasion through the hypodermis into the dermal-epidermal junction. Inset: Distinct tumor emboli are visible within the dermal layer of the skin in the H&E section of FC-IBC01 xenograft tissue (inset). C. Confocal microscopy combined with triple color immunofluorescence staining demonstrates that mice bearing FC-IBC01 tumors form tumor emboli within the dermis that express E-cadherin (green fluorescence) and are enwrapped by lymphatic vessels, identified by specific staining for podoplanin (red fluorescence). The DNA dye Topro-3 (blue fluorescence) identifies nuclei. D. FC-IBC01 tumor emboli contain ALK protein (green fluorescence) and are encircled by podoplanin stained lymphatic endothelium (red fluorescence). E. Dose response analysis of tumors cells freshly isolated from the patient designated as FC-IBC01 demonstrating response to Crizotinib and resistance to Paclitaxel.