S1P inhibits invasion and affects expression, secretion and activity of MMP2 and MMP9 in C643 cells.

<p>(A) Cells were treated with S1P (100 nM for 2 h, 4 h 6 h and 8 h). The representative western blots show the expression of MMP2 and MMP9. β-Actin was used as loading control and the normalized data are presented in the graphs. (B) Cells were treated with S1P (100 nM for 2 h, 4 h, 6 h and 8 h). MMP2 and MMP9 mRNA expression was measured with quantitative real-time PCR. (C) Cells were treated with S1P (100 nM for 2 h, 4 h, 6 h and 8 h). The representative western blots show the secretion of MMP2 and MMP9. (D) Cells were treated with S1P (100 nM for 2 h, 4 h, 6 h and 8 h). The representative zymography blot shows the activity of MMP2 and MMP9. The data were normalized to the protein content on the plates and are presented in respective graphs. (E) C643 cells were allowed to invade through collagen IV towards 5% lipid-stripped FBS in the presence or absence of S1P (100 nM for 2 h, 4 h, 6 h and 8 h). Representative images of invasion inserts are shown. The scale bar is 50 μm. The normalized data in the graphs are the mean ± S.E.M, n = 3–6. Asterisks (*) denote the statistically significant differences compared with respective control. Data were analyzed with one-way ANOVA and Bonferroni’s post hoc test (* <i>P</i> < 0.05; ** <i>P</i> < 0.01; *** <i>P</i> <0.001).