Single cell nuclei were tracked over time, and migratory speed was calculated by dividing the length of the migration path by the total movie time.
This LFA-1 links to the cytoskeleton through α-actinin-1 and disruption at the level of integrin or actin results in loss of cell spreading and migratory speed due to a failure of attachment at the leading edge.
For the wound-healing assay, cultured cells were wounded, photographed each hour, and quantified as the average linear migratory speed of the wound edges over 12∼16 hours.
In addition to G αq, CCR7 also used another pathway involving Rho/Pyk2/cofilin and presumably depended on G12/G13 to control the migratory speed of DCs [ 81].
Consequently, over 64% of APC pseudopods in the laser-treated or laser+OVA-treated groups had a migratory speed above 0.5 µm/min, whereas less than 10% of pseudopods in control and OVA alone groups showed a migratory speed above 0.5 µm/min (Figure 2D).
Likewise, the cell pseudopod migratory speed as measured by a traveling distance per min was faster with laser illumination irrespective of whether or not OVA was administered (0.71±0.08 vs. 0.23±0.03 µm/min in the absence of OVA and 0.76±0.08 vs. 0.31±0.03 µm/min in the presence of OVA, p<0.001) (Figure 2D).
Migratory speed was not reduced in EphA2/EphA4 siRNA-treated cells compared to control siRNA-treated cells (Fig. 1E).
Quantification revealed that in deep cortical layers the migratory speed of radially migrating 5-HT6R-shRNA1 TOM+ cells was significantly decreased versus scram-shRNA (Fig. 3D; supplementary material Movies 1 and 2).
The migratory speed of EA.hy926 cells treated with culture supernatants of Corin- siRNA H9c2 cardiomyoblasts was markedly decreased compared with that in NC- siRNA group using the wound healing assay (Fig. 6a).
Further, we found that the highly invasive MDA-MB-231 cells are able to produce a certain amount of HGF, TGF- β or bFGF, which slightly influenced their migratory speed.
