Fig. 8: Ligand-bound integrins are activated to the EO state at lower force than required for spreading on RGD.
From: Single-molecule characterization of subtype-specific β1 integrin mechanics
a Schematic of immobilization of biotin-functionalized cells for activated integrin β1 detection. Cholesterol (CLS) linked to biotin with PEG (~13 nm) immobilizes cells on neutravidin and TGT-coated surfaces. b Cell images and AF647-12G10 Fab (20 nM) signal 1 h after seeding. Scale bar, 10 µm. c–e Cell area and 12G10 Fab signal were measured 30 min after seeding (n = 43, 55, 62, 69, 21, 29; 83, 59, 83, 92, 68, 62 cells from two independent experiments). c Close-contact enclosed area of cells. Cells did not spread on the control or 12–33 pN RGD-TGT surfaces. d The number of 12G10 Fab counted per cell. Fluorescent Fab signal was calibrated by analyzing stepwise signal increase due to nonspecific Fab binding to surfaces outside the area imaged with cells (Supplementary Fig. 4a). e Density of 12G10 Fab for each cell. Lines show mean ± SE. Two-sided t-test for p-values. Not significant (ns), p > 0.05. Source data are provided as a Source Data file.
