fig1
Figure 1. Flow chart describing the experimental in vivo model system. All the procedures have been performed according to Directive 2010/63/EU and Italian Decree Law 26/2014. They were approved by the EU Research Executive Agency, the Intramural Regina Elena Board for Animal Welfare, and the Italian Ministry of Health (700-2015-PR, dated July 17, 2015). Tumor xenotransplants were established by inoculating 3 × 106 cells from the HT-29 and LoVo cell lines in the flank of 4-month old Nu/CD1 mice (Charles River Laboratories, Italy). Xenotransplants were allowed to grow to two different sizes (300 and 1000 mm3, 6 mice per group for each of the two cell lines). Tumors were taken at sacrifice along with blood. Frozen tissues were used as the source of miRNAs. Blood was collected in 6 mL BD Vacutainer K2E tubes (BD, 368857) and centrifuged within 1 h at 2000 ×g for 20 min at 4 °C. Plasma was recovered and further centrifuged at 16,000 ×g for 10 min at 4 °C to remove cell debris, and stored at -80 °C until extraction
