Figure 3. The FN synergy site is required to establish tensioned FN-α5β1 bonds.

(A) Quantification of adhesion strength. 7 × 10⁵Fn1-KO cells attached onto purified, full-length (fl) pFN^wt or pFN^syn or FNIII7-10^wt or FNIII7-10^syn and spun with a spinning disk device (n = 7 independent experiments with fl-FN; n = 3 independent experiments with FNIII7-10; mean ± sem). (B) Western-blot analysis (left) and quantification (right) of cross-linked α5 integrins to pFN^wt or pFN^syn before and after applying shear forces (n = 6 independent experiments; mean ± sem). (C) Western-blot analysis (left) and quantification (right) of pY397- and pY861-FAK levels in Fn1-KO cells plated on pFN^wt or pFN^syn (n = 6 independent experiments; mean ± sem). Statistical significances were calculated using the Student t-test; *p<0.05, **p<0.01 and ***p<0.001.

DOI: http://dx.doi.org/10.7554/eLife.22264.010

Figure 3—figure supplement 1. Representative spinning disk experiment showing the cell distribution profile against the shear force.

The shear force was calculated according to the method described by Friedland and Boettiger (Friedland et al., 2009). The force corresponding to the inflexion point of the curve is the τ50.

DOI: http://dx.doi.org/10.7554/eLife.22264.011