Additional file 8: of ETV4 transcription factor and MMP13 metalloprotease are interplaying actors of breast tumorigenesis

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

Additional file 7: of ETV4 transcription factor and MMP13 metalloprotease are interplaying actors of breast tumorigenesis

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

Additional file 10: of ETV4 transcription factor and MMP13 metalloprotease are interplaying actors of breast tumorigenesis

Figure S8. Metastasis-free survival analysis from the publicly available NKI datasets of breast tumors. a Metastasis-free survival (MFS) curves for patients with breast tumors according to Low-ETV4 (n = 12), High-ETV4 and Low-MMP13 (n = 243), or High-ETV4 and High-MMP13 (n = 9) mRNA levels. ****P ≤ 0.0001. b Metastasis-free survival (MFS) curves for breast tumor patients according to Low-MMP13 (n = 255) or High-MMP13 (n = 9) mRNA levels. ****P ≤ 0.0001. c Metastasis-free survival (MFS) curves for patients with breast tumors according to Low-ETV4 (n = 13) and High-ETV4 (n = 251) mRNA levels. ****P ≤ 0.0001. (PDF 19 kb

Additional file 6: of ETV4 transcription factor and MMP13 metalloprotease are interplaying actors of breast tumorigenesis

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

Additional file 2: of ETV4 transcription factor and MMP13 metalloprotease are interplaying actors of breast tumorigenesis

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

Additional file 11: of ETV4 transcription factor and MMP13 metalloprotease are interplaying actors of breast tumorigenesis

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