Fig. 6.

Expression of α-Klotho, vascular endothelial growth factor (VEGF)₁₆₅b, VEGF receptor-2 (VEGFR-2) and transient receptor potential canonical-1 (TRPC-1) in dermal microvascular endothelial cells from healthy controls (H-MVECs) and patients with systemic sclerosis (SSc) (SSc-MVECs). a, b Representative microphotographs of immunocytochemical staining for α-Klotho in H-MVECs (a) and SSc-MVECs (b) detected with biotinylated secondary antibodies and streptavidin peroxidase. c-f Protein lysates from H-MVECs and SSc-MVECs were subjected to western blotting analysis using anti-α-Klotho (c), anti-VEGF₁₆₅b (d), anti-VEGFR-2 (e) and anti-TRPC-1 (f) antibodies. 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