Migration of cells treated with Gal-3, Gal-3C and LNnT.

<p>Cells were treated for 21 h with buffer control, Cytochalasin D (197 nM), Gal-3 (50 μg mL^-1), Gal-3C (50 μg mL^-1), and LNnT (100 mM). Migration was normalized and compared to buffer control; *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.005; ****, p ≤ 0.0001.</p

Viability of cells under cell migration conditions.

<p>Viability of cells under cell migration conditions.</p

Clustering of integrins is increased on Gal-3 treated cells.

<p>Cells were stained using the same anti-α5-Cy5 conjugate employed for tracking experiments. Ten fields of stained cells were analyzed using ImageJ to identify clusters and measure their size. Treatment with Gal-3 resulted in an increase in the size of integrin clusters. See Table B and Figure A in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184378#pone.0184378.s001" target="_blank">S1 File</a>. Data were compared to a PBS control, or PBS containing BME (control) using a student’s t-test to determine p values; *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.005; ****, p ≤ 0.0001.</p

Model of Gal-3 interactions with integrin.

<p>(<b>a.</b>) Glycosylated receptors, such as the integrins, will have reduced binding sites for Gal-3 if they are heavily sialylated. (<b>b.</b>) Removal of sialic acids by neuraminidase enzymes (or decreased SiaT activity) will increase the number of Gal-3 binding sites present, and should increase oligomerization (only a dimer is shown for clarity). Oligomers likely interact with cytoskeletal regulators, including talin,[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184378#pone.0184378.ref089" target="_blank">89</a>] which lead to increased mobility through active processes. (<b>c.</b>) Addition of exogenous Gal-3C or a competitive binder (e.g. LNnT) will disrupt the formation of oligomers. This will occur either by (<b>d.</b>) competition for dimerization sites or (<b>e.</b>) blocking dimer binding sites.</p

Lateral mobility of integrins.

<p>Lateral mobility of integrins.</p

Lateral mobility of integrins is modulated by the presence of saccharides and lectins.

<p>The lateral mobility of integrins were determined using SPT, and the data from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184378#pone.0184378.t001" target="_blank">Table 1</a> are shown. Each sample population is shown as a bean plot,[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184378#pone.0184378.ref087" target="_blank">87</a>] with the logarithmic median of the diffusion coefficients indicated by a solid line for each population.[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184378#pone.0184378.ref087" target="_blank">87</a>] Each population is shown with a density estimate and horizontal lines indicate individual diffusion coefficient measurements. Gal-3C and Gal-3 treatments are shown for 50 μg mL^-1 concentrations. Diffusion coefficients are given as log(D), where D is in units of x 10⁻¹⁰ [cm²s^-1] or x 10⁻² [μm²s^-1]. Data were compared to a PBS control using a student’s t-test to determine p values; *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.005; ****, p ≤ 0.0001.</p

Viability of cells treated with LNnT, Gal-3C, and Gal-3.

<p>Cells were treated for 21 h with buffer control, Gal-3 (50 μg mL^-1), Gal-3C (50 μg mL^-1), and LNnT (100 mM). Viability of each condition were measured using MTS assay.[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184378#pone.0184378.ref088" target="_blank">88</a>] Viability for each condition was normalized and compared to buffer control; *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.005; ****, p ≤ 0.0001.</p