Figure 3

Relative mRNA and protein expression levels of MMPs and MMPs inhibitors in the MDA-MB-231 cell line treated with different concentrations of TGF-β1. The mRNA expression levels of MMPs (MMP-2, MMP-9 and MMP-14) and MMPs inhibitors (TIMP-1, TIMP-2, TIMP-3 and RECK) were analyzed by qRT-PCR using the total RNA from the MDA-MB-231 cells treated with 0, 1, 5 or 10 ng/mL TGF-β1 for 20 h. The levels of pro-enzyme and active MMP-2 and MMP-9 proteins were evaluated by zymography. Total protein lysates were used to measure the protein expression levels of MMP-14 and RECK by Western Blotting. The β-tubulin levels were used as protein loading control for Western Blot analysis. Conditioned medium was used to analyze the protein levels of TIMPs (TIMP-1, TIMP-2 and TIMP-3) by Western blotting. The results are presented in graphics as means ± standard errors from three independent experiments. The Western-blotting figures are representative of one experiment. *, p < 0.05, **, p < 0.01 and *** p < 0.001, all versus control (untreated cell).