Fig. 6

Effects of reduction of TIMP-2 mRNA on proliferation of FT282, JHOS2 and OVCAR4 cell lines. T2-KD is representative of a pool of all three TIMP-2 siRNAs. P are parental cells without siRNA treatment. Cont are parental cells transfected with scrambled siRNA. The experiments were performed 48 h after TIMP-2 knockdown by siRNA. a MTT assay was performed as described in Methods. Bar graphs represent mean ± SEM of 3 different MTT experiments. Significance was obtained using One-way ANOVA. *p < 0.05, ***p < 0.001, ****p < 0.0001. b Cells were stained for EdU and analysed as described in the Methods. Results are expressed as % of EdU positive cells (or cells in the S-phase of cell cycle progression). Bar graph represent n = 3 for each experiment. c mRNA expression of CDC25B; d CDC25C and e CDC25A were evaluated by qRT-PCR. Statistical significances were obtained using One-way ANOVA and are indicated by *p < 0.05, **p < 0.01, ***p < 0.001. f Effect of reduction in TIMP-2 mRNA on invasion of FT282, JHOS2 and OVCAR4 cell lines. T2-KD (blue line) is a representative of a pool of all three TIMP-2 siRNAs. Cont are parental cells transfected with scrambled siRNA (pink line). Invasion assays were assessed by xCELLigence real-time cell analysis. For assessment of invasion, the electrodes were coated with Matrigel and the bottom of the well contained reduced serum medium (OPTIMEM). Significance was assessed by linear regression analysis of two slopes; ****p < 0.0001