Fig. 5

Differential activation of Plexin-D1 (PlxnD1)/semaphorin 3E (Sema3E) cell signaling pathway in dermal microvascular endothelial cells (MVECs) from healthy controls (H) and patients with systemic sclerosis (SSc). Protein lysates from H-MVECs at basal condition and after stimulation with recombinant human vascular endothelial growth factor (VEGF)-A165, sera from healthy controls and patients with SSc (both n = 5), and from SSc-MVECs at basal condition were assayed with antibodies recognizing (a) the activated form of PlxnD1 (phosphorylated in C-ter domain, a.a. 1635–1647), (b) the N-terminal Sema domain region of PlxnD1, (c) Sema3E N-terminal region, and (d) PlxnD1 C-terminal domain. Representative immunoblots are shown. The densitometric analysis of the bands normalized to α-tubulin is reported in the histograms. Data are mean ± SD of optical density (OD) in arbitrary units. Student’s t test was used for statistical analysis; p values are indicated in each panel. Results are representative of three independent experiments performed with each one of the five H-MVEC and five SSc-MVEC lines