Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2015 Oct 12;211(1):173–190. doi: 10.1083/jcb.201505007

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2015 Fiore et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

PMC Copyright notice

Figure 4. — Thy-1 binds inactive α_vβ₃ and is required for membrane raft recruitment to FAs, mechanosignaling, and rigidity sensing. (a) α_v integrin was immunoprecipitated from human NLFs in suspension versus spread on FN for 30 min or overnight, and α_v and Thy-1 were immunoblotted. (b) Micrographs of PLA between α_vβ₃ (mAb 23C6) and Thy-1, pxn immunostaining, and F-actin in NLFs treated with nonbinding control talin (1–405)^A/E (cont. vector), wild-type THD (1–405), or 2 mM Mn²⁺ integrin activating conditions. Quantification of PLA cell surface density (mean ± SEM). Bar, 20 µm. (c) PLA signals were masked with pxn immunostaining and the surface density of PLA was segregated into inside versus outside FAs regions and quantified. Bar, 20 µm. (d) Quantification of PLA cell surface density in CHO.B2(hα_vβ₃) expressing human Thy-1^WT, Thy-1^/CD8, and Thy-1^D37E. PLA puncta, mCherry (transfection control), and cell nuclei are shown. Bar, 20 µm. (e) Immunoblots of proteins associated with adhesion complex or total lysate before (− force) or after (+ force) 3 min force application in Thy-1^WT and Thy-1^D37E RFL-6 cells. Averages of n = 3 independent experiments for association with FN-bead adhesion complexes before (− force) or after 3 min force application (+ force) were quantified. Significance was calculated between all groups for individual adhesion components. (f) Activation of RhoA after magnetic force stimulation of FN beads for the indicated time periods in Thy-1^D37E RFL-6 cells. Apparent rate constant (K) for a single-phase association model is indicated; plateau phase of Thy-1^D37E was statistically different from Thy-1^WT (n = 5; unpaired t test, P < 0.05). Immunostaining for FAs and F-actin (g), single-cell cortical stiffness measurements (h), and FA size (i) of Thy-1^D37E RFL-6 cells on FN-PA substrates of varying stiffness. Bar, 50 µm. Statistical significance was calculated using Kruskal-Wallis nonparametric test with Dunn’s multiple comparison. All other statistical significance was calculated using one-way analysis of variance and Tukey’s post test. *, P < 0.05; **, P < 0.01; ***, P < 0.001 between indicated groups.