Extended Data Figure 9: Effect of warfarin on leakiness, metastasis and clonal abundance.

a, Levels of prothrombin fragments F1 and F2 (active component of blood coagulation), as quantified by ELISA, after administration of warfarin in the drinking water of mice (P < 0.01, Wilcoxon rank-sum, n = 4 mice). b, DAPI and dextran Alexa 647 staining to visualize vascular leakage in primary tumours derived from the pool of 23 clonal lines. Animals were administered drinking water either with or without 10 mg ml⁻¹warfarin. c, Leakiness index of primary tumours resulting from orthotopic injection of the pool of 23 clonal lines. After injection mice were administered drinking water with no warfarin or water containing 10 mg ml⁻¹ warfarin (P < 0.000005, Wilcoxon rank-sum, n = 10 mice). d, Counts of lung metastatic nodules in animals that were injected intravenously with the pool of 23 clonal lines administered drinking water with no warfarin or water containing 10 mg ml⁻¹ of warfarin (P < 0.05, Wilcoxon rank-sum, n = 10 mice). e, The pool of 23 clonal lines was injected orthotopically and the mice administered drinking water with no warfarin or 10 mg ml⁻¹ warfarin. The percentage abundance of each clone in the cardiovascular CTCs of animals (either pre- or post-injection, P < 0.002, n = 7 mice for no warfarin, n = 7 mice for pre-injection and n = 5 mice for post-injection). f, The clone proportions in the resultant primary tumours and lung metastases described in (e) (P < 0.003, Wilcoxon rank-sum, for lung metastases, n = 7 mice for no warfarin, n = 7 mice for pre-injection and n = 5 mice for post-injection). For all box plots, the edges of the box are the twenty-fifth and seventy-fifth percentiles. The error bars extend to the values q3 + w(q3 − q1), and q1 − w(q3 − q1), in which w is 1.5 and q1 and q3 are the twenty-fifth and seventy-fifth percentiles.