Supplementary Figure 7: E-selectin binding to tumour cells induces MET.

(a) Confocal imaging of bone metastasis of Nu/Nu mice after intra-iliac artery injection of GFP-labelled SCP28 cells. Mice were labelled with a retro-orbital injection of 10 μg anti-CD31 prior to euthanasia. Scale bars represent 50 μm. (b) Confocal imaging of BM2-GFP bone lesions in Nu/Nu mice stained for E-selectin and Glg1. Scale bars represent 100 μm (c) Confocal imaging of E-selectin-induced intensity changes in Keratin-14 expression in BM2 cells after culturing on E-selectin compared to IgG-coated plates (10 μg/mL) for 24h. Scale bars represent 20 μm. Data representative of 3 independent experiments (a-c). (d, e) GSEA of multiple EMT-related signatures, including Blick EMT¹ (d), Claudin-low (GSE18229), Sarrio-EMT (GSE8430), and Luminal (GSE22446) (e), in the ranked gene list of BM2 cells cultured in E-selectin vs. IgG-coated plates. Single mRNA isolation used for each condition in each cell line. p and q statistics by GSEA software, n = 108 gene sets queried. (f) Western blot of N-cadherin and Slug from M1a cells bound to IgG or E-selectin-coated plates (10 μg/mL) for 24h. Data representative of 2 independent experiments. (g, h)Confocal imaging of bone lesions from BM2-GFP injected mice stained for E-cadherin (g) and EpCam (h). Scale bars represent 100 μm (g) and 50 μm (h). Data representative of >5 independent biological replicates. Unprocessed original scans of the blots in f are shown Supplementary Fig. 9.