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. 2023 Jan 10;13:1085892. doi: 10.3389/fphar.2022.1085892

FIGURE 2.

FIGURE 2

Truncated sema3A is unable to induce NRP-1 mediated signaling in endothelial cells (A). Cell contraction assay was performed on HUVEC cells in the presence of 200 ng/mL of purified P-sema3A or T-sema3A or an appropriate amount of a control elution buffer for 30 min at 37°C. After the incubation time, the cells were photographed using a phase-contrast inverted microscope: A round shape is the natural morphology of the HUVEC (black arrow), while as a result of sema3A, the cytoskeleton collapsed, and the cells lost their round shape (red arrows). The quantification graphs represent the percentage of contracted cells from total cells (N = 8 independent experiments). The results were analyzed using Kaluza software, and the statistical significance was calculated using one-way ANOVA and Kruskal–Wallis test (**** = p-value < 0.0001). (B). Quantitative real-time contraction assay was performed on HUVEC cells placed in an xCELLigence real-time cell analyzer. At time 0, 200 ng/mL of purified P-sema3A or T-sema3A or an appropriate amount of a control elution buffer was added to the wells and the electrical impedance through the E-plates electrodes (which displayed as Cell Index (CI)) was measured for 30 min.