Figure 4
Coronin 1C null cells degrade gelatin more efficiently than Coronin 1C-expressing cells. (a) Null, Rescue and OE cells were plated on FITC-conjugated 2D gelatin matrices for 24 h, fixed, and stained for F-actin with phalloidin-568. Black spots in the gel matrix are indicative of degradation, and degradative spots of invadopodia that colocalize with actin puncta are shown with red arrows. Scale bars = 25 µm. Zoom = 3.5X. (b) Percent of cells that contain degraded spots of the matrix beneath the cell body. (c) Number of degraded spots per cell showing degradation. (d) Number of degraded spots per area of 2D matrix uninhabited by a cell body. N cells B-C > 30 from 3 biological replicates. N of images analyzed for D = 30 from 3 biological replicates. (e) Mean velocities ± 95% CI of Null, Rescue and OE cells migrating on 2D gelatin matrices over 16 h. N cells > 40 from 3 biological replicates. (f) Null, Rescue, and OE cells were plated on FN-coated glass coverslips, fixed, and surface labeled with MT1-MMP. Integrated fluorescence density was calculated for individual cells and graphed as mean + /− SEM. N > 70 for each cell line over 3 biological replicates. ** P < 0.01, *** = P < 0.001.
