Journal: Nature communications

Article Title: Fluid shear stress activates YAP1 to promote cancer cell motility.

doi: 10.1038/ncomms14122

Figure Lengend Snippet: Figure 1 | Fluid flow stimulates motility and matrix metalloprotease activity. (a) Cylindrical PDMS fluidics channels coated in collagen support a monolayer of cells. Flow of media through the culture chamber exposes cells to WSS of 0.05 dyne cm 2. Scale bar on bright field photomicrograph of PC3 cells within the scaffold represents 400 mm. (b) Filopodia formation in response to WSS is extensive. Scale bar in left panel, 10 mm, scale bar in right panel, 5 mm. (c) Transcription of MMP2 and MMP9 is stimulated by WSS (n ¼ 3 independent experiments; Kruskal–Wallis one-way ANOVA, Po0.001). (d) Total MMP activity measured by fluorogenic peptide substrate digestion assays was increased by exposure to 6 h WSS (n ¼ 3 independent experiments; unpaired t-test, **Po0.0001). (e) Spatial tracking of PC3 and DU145 cancer cell movement during 6 h of time-lapse imaging, where each cell lies at the origin (0,0) at t ¼ 0 h. Plots depict motility of individual cells in one representative experiment. (f) Quantification of migration speed reveals increased cellular velocities of individual cells under WSS. (n ¼ 7 independent experiments, two-tailed t-test, **P ¼ 4.22E 18 for PC3 cells; n ¼ 3 independent experiments, two-tailed t-test, **P ¼ 1.56E 9 for DU145 cells). (g) Time segmented migration speed of cells after WSS initiation (Kruskal–Wallis one-way ANOVA, *Po0.05, **Po0.01). Error bars represent±s.e.m.

Article Snippet: Total MMP fluorogenic peptide substrate (Mca-K-P-L-G-L-Dpa-A-R-NH2, R&D system) was added to a 10mM final concentration and incubated for 1 h at room temperature.

Techniques: Activity Assay, Imaging, Migration, Two Tailed Test