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Fig. 5 | Breast Cancer Research

Fig. 5

From: ETV4 transcription factor and MMP13 metalloprotease are interplaying actors of breast tumorigenesis

Fig. 5

MMP13 acts as a regulator of ETV4 tumorigenic-induced response in mammary epithelial MMT cells. a Relative MMP13 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-ETV4 + shCtrl cells was arbitrarily = 1. Error bars indicate SD. ****P ≤ 0.0001. b Zymographic analysis of MMP13 protein activity (55 kDa) from the supernatant of MMT-ETV4 + shCtrl and MMT-ETV4 + shMMP13 cells. c MMT-ETV4 + shCtrl and MMT-ETV4 + shMMP13 cell proliferation analysis by cell counting. The two charts represent the number of counted cells at 10, 35, 55, 80, and 100 hours. Experiments were conducted three times in triplicate. Error bars indicate SD. ****P ≤ 0.0001; ***P ≤ 0.001; **P ≤ 0.01. d MMT-ETV4 + shCtrl and MMT-ETV4 + shMMP13 cell migration analysis using a Boyden chamber culture system. Histogram represents the relative number of counted cells that migrated to the lower side. The number of MMT-ETV4 + shCtrl cells was arbitrarily = 1. Experiments were conducted three times in triplicate. Error bars indicate SD. ****P ≤ 0.0001. The lower panel depicts a representative picture of each experiment. Scale bar = 100 μm. e MMT-ETV4 + shCtrl and MMT-ETV4 + shMMP13 cell invasion analysis using a Boyden chamber culture system coated with Matrigel®. Histograms represent the relative number of counted cells that invaded to the lower side. The number of MMT-ETV4 + shCtrl cells was arbitrarily = 1. Experiments were conducted three times in triplicate. Error bars indicate SD. ****P ≤ 0.0001. The lower panel depicts a representative picture of each experiment. Scale bar = 100 μm. f Anchorage-independent growth. MMT-ETV4 + shCtrl and MMT-ETV4 + shMMP13 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. The lower panel depicts a representative picture of each experiment. Scale bar = 100 μm

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