Figure 5
Impact of KLK12 on the migration of lung-derived endothelial cells: (A) Endothelial cell migration was analyzed in a Boyden chamber assay after treatment with 500 nM FN in the presence or absence of 10 nM KLK12. After 24 h, migrated cells were stained with DAPI and counted. (B) Endothelial cell migration was analyzed by time lapse video microscopy after treatment with 10 nM KLK12 in the presence or absence of 500 nM FN (5 h at 37 °C, 5% CO2). At least 40 cells per condition were analyzed on a total of three biological replicates. Cells were tracked (using Imaris software) on the basis of nuclear staining and bright field images, and the displacement length (left) and velocity (right) were determined. (C) Endothelial cell migration after treatment with 10 nM KLK12 in the presence or absence of 500 nM of isotype control antibody or pAbFN-KLK12 targeting the KLK12-cleavage site on FN, according to the protocol described in B. *p < 0.05, **p < 0.01, ***p < 0.001 in a two-way Kruskal-Wallis analysis of variance.
