Figure 5:

Fibronectin is enriched around breast cancer spheroids in high stiffness scaffolds due to Mena-mediated assembly. A.) Heatmaps of immunofluorescent staining of fibronectin demonstrated the development of a high intensity halo around spheroids in 12 kPa gels. The halo increased in spread and intensity over time, with clear differences between low and high stiffness conditions. Scale bar=150 μm, solid line is mean and shaded area of line plot is standard deviation, n=3 line intensities per spheroid and 3 spheroids per condition. B.) Heatmaps of fibronectin staining at 24 hr post embedding of spheroids treated with either scrambled (left) or MENA-targeted (right) siRNA demonstrated the knockdown of MENA decreased the intensity and spread of fibronectin signal. C.) Similarly, fibronectin heatmaps at 24 hr post embedding of spheroids treated with an anti-α5 antibody (right) had reduced intensity and spread of the halo compared to untreated controls (left). Scale bar=150 μm.