Figure 2

Decreased expression of epidermal growth factor-like domain 7 (EGFL7) in the affected skin and dermal microvascular endothelial cells (MVEC) of patients with systemic sclerosis (SSc). (A-F) Representative microphotographs of skin sections from healthy controls (A-C; n = 10) and SSc patients (D-F; n = 16) immunostained for EGFL7 (red) and counterstained with 4′,6-diamidino-2-phenylindole (DAPI; blue) for nuclei. Arrows indicate microvessels. (G and H) Representative microphotographs of skin sections from healthy controls (G) and SSc patients (H) double immunostained for EGFL7 (red) and the pan-endothelial cell marker CD31 (green) and counterstained with DAPI (blue). Original magnification and scale bars are indicated in each panel. (I) Densitometric analysis of EGFL7 immunofluorescent staining in dermal microvessels expressed as optical density in arbitrary units (a.u.). Data are shown as box plots. Each box represents the 25th to 75th percentiles. Lines inside the boxes represent the median. Lines outside the boxes represent the 10th and the 90th percentiles. Mann–Whitney U-test was used for statistical analysis. (J) Western blotting of protein lysates from control dermal MVEC (n = 3) and SSc MVEC (n = 3) analyzed using anti-EGFL7 antibodies. Representative immunoblots are shown. The densitometric analysis of the bands normalized to α-tubulin is reported in the histograms. Data are mean ± standard error of the mean of optical density in a.u. Student t-test was used for statistical analysis.