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Figure S5. Expression of ETV4 in MMT-shMMP13-repressing cells. a Relative ETV4 mRNA expression in the MMT-ETV4 + shCtrl and MMT-ETV4 + shMMP13 cells determined by real-time PCR and normalized to cyclophilin A levels. mRNA expression in MMT-Ctrl + shCtrl cells was arbitrarily = 1. Error bars indicate SD. The results were not statistically significant. b Western blot analysis of ETV4 protein expression (61 kDa) in the MMT-ETV4 + shCtrl and MMT-ETV4 + shMMP13 cells. GAPDH expression served as the loading control. (PDF 71 kb)

Figure S4. Expression of MMP13 in MMT cells overexpressing or repressing MMP13. a and b Relative MMP13 mRNA expression in the MMT-Ctrl and MMT-MMP13 (a) or MMT-shCtrl and MMT-shMMP13 cells (b) determined by real-time PCR and normalized to cyclophilin A levels. mRNA expression in MMT-Ctrl cells was arbitrarily = 1. Error bars indicate SD. ****P ≤ 0.0001. c Western blot analysis of MMP13 protein expression (60 kDa) in the MMT-Ctrl and MMT-MMP13 cells. GAPDH expression served as the loading control. d Zymographic analysis of MMP13 protein activity (55 kDa) from the supernatant of MMT-Ctrl and MMT-MMP13 cells. (PDF 72 kb)

Figure S3. ChIP experiment for ETV4 and MMP13 in MMT and TAC cells. PCR detection of the MMP13 promoter region after ETV4 immunoprecipitation in MMT-ETV4 (left panel) and TAC-ETV4 (right panel). Primers allowing the amplification of the proximal MMP13 promoter region containing EBS are schematized in the lower panel of Fig. 2. Cyclin D2 was used as a positive control [8]. Immunoprecipitation with a nonrelevant antibody (IgG) was used as negative control. (PDF 60 kb)

Table S1. Pathological and clinical characteristics of patients in relation to metastasis-free survival (MFS). (PDF 33 kb)

Table S2. Multivariate Cox proportional hazards analysis of MFS for MMP13 and ETV4 expression levels in the series of 456 breast tumors. (PDF 42 kb)

Figure S6. The repression of MMP13 reduces the anchorage-independent growth capacity of MMT-ETV4-overexpressing cells. a Relative MMP13 mRNA expression in the transiently transfected MMT-siCtrl and MMT-siMMP13 cells determined by real-time PCR and normalized to cyclophilin A levels. mRNA expression in MMT-siCtrl cells was arbitrarily = 1. Error bars indicate SD. ****P ≤ 0.0001. b Anchorage-independent growth. MMT-ETV4-siCtrl and MMT-ETV4-siMMP13 cells were cultured for 10 days in soft agar. This histogram represents the number of clones counted for experimental time points. Soft agar assays were conducted three times in triplicate. Magnification × 5. Error bars indicate SD. ****P ≤ 0.0001. (PDF 45 kb)

Figure S2. ETS and AP-1 binding sites are highly conserved among mouse, human, and rabbit. Nucleotide sequence comparison of mouse, human, and rat proximal MMP13 promoters. Shaded boxes indicate the conserved ETS and AP-1 binding site sequences. (PDF 46 kb)

Figure S1. Validation of the overexpression of ETV4 and MMP13 in MCF10A cells. a and b Relative ETV4 mRNA (a) and MMP13 mRNA (b) expression in the MCF10A-Ctrl and MCF10A-ETV4 cells determined by real-time PCR and normalized to cyclophilin A levels. mRNA expression in MCF10A-Ctrl cells was arbitrarily = 1. Error bars indicate SD. *P ≤ 0.1. c and d Western blot analysis of ETV4 protein expression (61 kDa) (c) and MMP13 protein expression (60 kDa) (d) in the MCF10A-Ctrl and MCF10A-ETV4 cells. GAPDH expression served as the loading control. (PDF 100 kb)

Figure S7. High MMP13 mRNA expression level is associated with a poor prognosis in breast cancer. a Metastasis-free survival (MFS) curves for patients with breast tumors according to Low-MMP13 (n = 321) or High-MMP13 (n = 135) mRNA levels. ***P ≤ 0.001. b Metastasis-free survival (MFS) curves for patients with breast tumors according to Low-ETV4 (n = 82) and High-ETV4 (n = 374) mRNA levels. The results were not statistically significant. (PDF 14 kb)