Fig. 2

Migration and invasion properties of HeLa clones. A, B 2D migration assay with trajectories (A) and migration speed (B). The migration trajectories of cells were computed from images recorded every 15 min for 1 h. C, D 3D migration assay. Trajectories along the x-y-z plane (C) and migration speed (D) of the WT and KD clones. E Activation status of Rac1 (Rac1-GTP) and PAK (phosphorylated PAK, pPAK) of the CTR, WT, and KD clones as compared to total Rac1 and PAK protein. F, G Collagen invasion assay. The morphological aspect of cells in type I collagen (F; scale bars: 100 μm) with individual cells magnified (inserts) and arrowheads pointing to membrane protrusions, as well as invasion index (G). Note that mutations of NDPK-D result in the invasion of HeLa cells into native type I collagen gel. Data show means ± SEM (n=3). *p< 0.05, ***p< 0.005 relative to control/empty vector (CTR); ^##p< 0.01 and ^###p< 0.005 relative to wild-type (WT). For clone abbreviations see Fig. 1; where indicated, two clones of the same type were analyzed