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. 2021 Mar 19;9:620128. doi: 10.3389/fbioe.2021.620128

FIGURE 7.

FIGURE 7

Matrix proteolysis is required for larger sprout diameters that are lumenized. (A) Representative image (individual z-slices) of collagen degradation along sprout stalk cells. F-actin (cyan), nuclei (magenta), collagen hybridization peptide (CHP; yellow). (B) Representative images (max intensity projections) of marimastat (MMS) treatment (1 μM) of sprouts cultured over 5 days with 250 nM sphingosine 1-phosphate (S1P) and 25 ng ml–1 phorbol 12-myristate 13-acetate (PMA) within 3 mg ml–1 collagen. (C) Representative images (single z-slice) of fluorescently labeled collagen (intensity heat map) with MMS treatment (1 μM) of sprouts cultured over 3 days with 250 nM S1P and 25 ng ml–1 PMA within 3 mg ml–1 collagen. (D,E) Quantifications of sprout diameter and collagen compaction from MMS treatment (1 μM). For sprout diameter: n ≥ 50 per condition and for collagen compaction: n ≥ 10 per condition. (F,G) Assessment of lumenization as a function of sprout diameter by parent vessel perfusion of 1 μm-diameter microsphere. Sample size for total sprouts analyzed for each group is indicated in bar plot. All data presented as mean ± SD; *indicates a statistically significant comparison with P < 0.05 (two-tailed Student’s t-test).