Figure 5. miR-335 regulates metastasis and invasion through suppression of SOX4 and TNC.

a, 5.0 × 10⁴ LM2 cells were transduced with short hairpin control vector, either of two shRNAs targeting SOX4, a hairpin targeting TNC, or an siRNA targeting TNC, and invasion of cells through a trans-well matrigel-coated membrane insert was quantified. n = 6; error bars represent s.e.m.; P-values based on a one-sided Student's t-test. b, 2 × 10⁵ LM2 cells transduced with either a control short hairpin, a short hairpin targeting SOX4 or one targeting TNC were inoculated intravenously into immunodeficient mice. Lung colonization was measured by bioluminescence and quantified. n = 7; error bars represent s.e.m.; P-values based on a one-tailed rank-sum test. c, Haematoxylin-and-eosin-stained images of representative lungs that were extracted at 4 weeks after cell inoculation. Representative images were obtained at 10× magnification. d, Kaplan-Meier curves for the combined Memorial Sloan Kettering and Erasmus Medical Center breast tumour cohorts (368 tumours) depicting metastasis-free survival of patients whose primary tumours expressed the miR-335 six-gene signature (positive) and those that did not (negative). n = 368; P-value based on the Mantel-Cox log-rank test.