Fig. 7

Cells expressing the phosphomimetic L-plastinS5E variant exhibit higher ability to degrade gelatin. a Representative images of the transduced MDA-MB-231 cells expressing the different GFP-fused L-plastin variants and degraded gelatin. Cells were plated on Alexa Fluor 568-labeled gelatin (red)-coated coverslips for 6 h and stained using Alexa Fluor 633-conjugated phalloidin (magenta) to visualize F-actin. GFP signal was amplified using the Alexa Fluor 488-conjugated GFP booster. Single confocal slices of the ventral surface of cells are shown. Areas of gelatin degradation can be identified as dark spots. Scale bar: 20 µm. b Quantification of cells associated with gelatin degradation. Results are expressed as means ± SEM of three independent experiments in which 100–150 cells per condition were assessed. One way ANOVA comparing all four groups showed no significance. c Quantification of the total degradation area normalized against total cell area. Results are expressed as means ± SEM of three independent experiments. One way ANOVA followed by Dunnett’s multiple comparison test relative to GFP transduced cells (*p < 0.05). d MMP activities measured by gelatin zymography. Conditioned media were collected after 24 h and equal amounts were loaded on the gel. Shown is a representative gel. The graph shows the densitometry of the MMP-9 degraded band relative to GFP control sample. Results are expressed as means ± SEM of three independent experiments. One way ANOVA followed by Dunnett’s multiple comparison test relative to GFP transduced cells (*p < 0.05)