Fig. 7

The migration and ECM degradation capacities of MV3 cells. a Histograms showing stable expression of LPL eGFP in MV3 cells. MV3 cells were transduced with lentiviral vectors expressing the indicated LPL constructs. Cells with intermediate LPL eGFP expression were sorted using FACS. b Quantification of the migration of MV3 cells expressing the indicated LPL constructs in 2D Transwell chambers and c in 3D invasion chambers. A total of 3 × 10⁴ cells were added to the upper chamber. Thirty minutes later, the cells were treated with the indicated concentrations of H₂O₂. The samples were fixed and mounted on microscopy slides. More than five xy focal planes per sample were imaged using LSM (×20 objective). d Representative confocal microscopy images showing the matrix degradation capacity of MV3 cells expressing eGFP alone or eGFP-tagged wt LPL, C101A LPL, or C42A LPL constructs. The cells were allowed to degrade Cy3-gelatin for 3 h in the presence of 25 µM H₂O₂. Shown are LPL eGFP (green), gelatin Cy3 (red) and merged images without (w/o) Cy3 (eGFP, F-actin (blue) and nuclei (white)). Scale bar = 20 μm; ×60 magnification. e Matrix degradation was quantified as the loss of fluorescence underneath the cells versus the total area of the cells. At least 150 cells were imaged per sample from three independent experiments. P-values were calculated by two-way ANOVA. The data are presented as mean ± SEM (n ≥ 3; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns nonsignificant)