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. 2019 Nov 4;4(1):bvz006. doi: 10.1210/jendso/bvz006

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© Endocrine Society 2019.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 8. — IGFBP-1 and angiogenic properties in vitro. (A) HUVECs were used to investigate the effects of rIGFBP-1 on endothelial cell tube formation on Matrigel. Treatment with rIGFBP-1 for 24 hours before being seeded onto Matrigel enhanced endothelial cell tube formation when compared to unstimulated cells (IGFBP-1 40 ± 4.26 V UN 26.75 ± 1.98). (B) HUVECs were used to investigate the effects of rIGFBP-1 on endothelial cell proliferation using an EDU incorporation assay. Treatment with rIGFBP-1 for 1hr before EdU was added enhanced endothelial cell proliferation when compared to unstimulated cells (IGFBP-1 15.24 ± 1.8 V UN 7.23 ± 1.67). (C) HUVECs were used to investigate the effects of rIGFBP-1 on endothelial cell sprouting using a cytodex bead assay. Representative images are shown. Beads were either grown in the presence of IGFBP-1 or not. (D) Beads grown in the presence of IGFBP-1 had a significantly increased number of sprouts (IGFBP-1 13.2 ± 0.79 V UN 10.7 ± 0.75). (E) Beads grown in the presence of IGFBP-1 had a trend for enhanced sprout length (IGFBP-1 2.7 ± 0.28 V UN 2.1 ± 0.23). (F) HUVECs were used to investigate the effects of mutant IGFBP-1 on endothelial cell tube formation on Matrigel. Treatment with WGD mutant IGFBP-1 for 24 hours before being seeded onto Matrigel had no effect on endothelial cell tube formation when compared with IGFBP-1 treated cells (IGFBP-1 37.2 ± 4.3 V WGD 30.6 ± 4.6). G: Treatment with amino-terminal deletion mutant IGFBP-1 for 24 hours before being seeded onto Matrigel reduced endothelial cell tube formation when compared with IGFBP-1 cells (IGFBP-1 37.18 ± 4. V NDEL 19.05 ± 1.3). (H) Treatment with an IGF-I neutralizing antibody and IGFBP-1 for 24 hours before being seeded onto Matrigel reduced endothelial cell tube formation when compared to IGFBP-1 only treated cells (IGFBP-1 69.3 ± 5.1. V IGF-1ab + IGFBP-1 38.7 ± 9.8). Data are presented as mean ± SEM. (*P ≤ .05; **P ≤ .01). N = 4 per group.