1. Nanoscale adhesive geometry modulates integrin–FN clustering.
Nanoscale adhesive geometry modulates integrin–FN clustering. (A) Integrin recruitment to Alexa-Fluor-555-labeled FN nanopatterns was analyzed using a cross-linking/extraction method that selectively retains FN-bound integrins. Fluorescence microscopy images for integrin binding (green) to FN (red) adhesive clusters with different geometrical configurations. Scale bars: 1 µm. (B) Frequency maps for integrin recruitment to adhesive zones generated by stacking individual images. The pseudo-color range represents the frequency of integrin localization to a spatial location with ‘warmer’ colors reflecting a higher frequency of localized integrin staining. (C) High magnification frequency maps for integrin recruitment to adhesive pads for the 45° and 90° orientations. (D) FN nanoislands in adhesive pads. Scale bars: 1 μm. (E) Binary images of the area corresponding to colocalization of integrin recruitment and FN nanoislands for the 45° and 90° pad orientation; images are oriented such that the edge closer to the center pad is at the top. (F) Frequency histograms of pad occupancy. Statistical analyses of frequency distributions revealed the following differences (P<0.05): 1000 nm×1>500 nm×4 = 333 nm×9>250 nm×4.
Sean R. Coyer et al. J Cell Sci 2012;125:
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