Figure 1
Figure 1. Human ProS inhibits Matrigel vascularization in vivo. (A) Matrigel plugs obtained from mice that were injected subcutaneously in the flank, with 0.4 mL of Matrigel alone (lane 1, control), Matrigel containing a mixture of proangiogenic factors (400 ng/mL bFGF, 400 ng/mL VEGF-A, and 50 units/mL heparin, plug 2) or human ProS at 25 μg/mL (plug 3) or a mixture of proangiogenic factors (bFGF at 400 ng/mL, VEGF-A at 400 ng/mL, heparin at 50 units/mL) supplemented with 25 μg/mL human ProS (plug 4). (B) Representative micrographs of hematoxylin and eosin-stained sections of Matrigel plugs under the experimental conditions; Matrigel alone (i, control), Matrigel containing a mixture of proangiogenic factors (ii, 400 ng/mL bFGF, 400 ng/mL VEGF-A, and 50 units/mL heparin) or human ProS at 25 μg/mL (iii) or a mixture of proangiogenic factors (bFGF at 400 ng/mL, VEGF-A at 400 ng/mL, and heparin at 50 units/mL) supplemented with 25 μg/mL ProS (iv). The arrows indicate vascular structures with lumens and red blood cells. (C) Quantification of vascular structures with lumens and red blood cells within Matrigel sections. (D) Representative micrographs of CD31 immunstaining of Matrigel plugs under the various experimental conditions; Matrigel alone (i, control), Matrigel containing a mixture of proangiogenic factors (ii, 400 ng/mL bFGF, 400 ng/mL VEGF-A, and 50 units/mL heparin) or human ProS at 25 μg/mL (iii) or a mixture of proangiogenic factors (bFGF at 400 ng/mL, VEGF-A at 400 ng/mL, and heparin at 50 units/mL) supplemental with 25 μg/mL ProS (iv). the arrows indicate blood vessel stained with CD31. Quantifications were performed from Matrigel plugs obtained from 3 independent experiments. Within each experiment and for each experimental condition, 3 individual plugs implanted in individual animals were analyzed. From each Matrigel plug, 3 sections were stained and analyzed. Results are expressed as percentage vascular area (C) or percentage CD31-positive area (E) ± SEM, using Fiji-win32 software. ***P < .001. **P < .01. *P < .05.

Human ProS inhibits Matrigel vascularization in vivo. (A) Matrigel plugs obtained from mice that were injected subcutaneously in the flank, with 0.4 mL of Matrigel alone (lane 1, control), Matrigel containing a mixture of proangiogenic factors (400 ng/mL bFGF, 400 ng/mL VEGF-A, and 50 units/mL heparin, plug 2) or human ProS at 25 μg/mL (plug 3) or a mixture of proangiogenic factors (bFGF at 400 ng/mL, VEGF-A at 400 ng/mL, heparin at 50 units/mL) supplemented with 25 μg/mL human ProS (plug 4). (B) Representative micrographs of hematoxylin and eosin-stained sections of Matrigel plugs under the experimental conditions; Matrigel alone (i, control), Matrigel containing a mixture of proangiogenic factors (ii, 400 ng/mL bFGF, 400 ng/mL VEGF-A, and 50 units/mL heparin) or human ProS at 25 μg/mL (iii) or a mixture of proangiogenic factors (bFGF at 400 ng/mL, VEGF-A at 400 ng/mL, and heparin at 50 units/mL) supplemented with 25 μg/mL ProS (iv). The arrows indicate vascular structures with lumens and red blood cells. (C) Quantification of vascular structures with lumens and red blood cells within Matrigel sections. (D) Representative micrographs of CD31 immunstaining of Matrigel plugs under the various experimental conditions; Matrigel alone (i, control), Matrigel containing a mixture of proangiogenic factors (ii, 400 ng/mL bFGF, 400 ng/mL VEGF-A, and 50 units/mL heparin) or human ProS at 25 μg/mL (iii) or a mixture of proangiogenic factors (bFGF at 400 ng/mL, VEGF-A at 400 ng/mL, and heparin at 50 units/mL) supplemental with 25 μg/mL ProS (iv). the arrows indicate blood vessel stained with CD31. Quantifications were performed from Matrigel plugs obtained from 3 independent experiments. Within each experiment and for each experimental condition, 3 individual plugs implanted in individual animals were analyzed. From each Matrigel plug, 3 sections were stained and analyzed. Results are expressed as percentage vascular area (C) or percentage CD31-positive area (E) ± SEM, using Fiji-win32 software. ***P < .001. **P < .01. *P < .05.

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