Figure 3.

THBS1 is pro-angiogenic in vitro. Monkey ovarian microvascular endothelial cells (mOMECs) were treated with human THBS1 protein at concentrations of 0.001–1 nM or no THBS1 (0 nM) as indicated. (A–C) Migration was assessed 24 h after plating on a porous membrane and treatment with THBS1. mOMECs (arrow) which migrated through pores (arrowhead) were stained with hematoxylin and eosin, photographed, and counted. Representative membranes from 0 nM (B) and 1 nM (C) treatment groups are shown. (D–F) Proliferation was assessed by Ki67 immunodetection in mOMECs cultured for 24 h with THBS1. Ki67 positive (arrows) and negative (arrowhead) cells are indicated in representative images from mOMECs treated with 0 nM (E) and 1 nM (F) THBS1. Data are expressed as a percentage of Ki67 positive cells among all cells counted. (G–M) Sprout formation in response to THBS1 treatment was determined after 1 day (G,I) and 2 days (H,J,L,M) in vitro. mOMECs coating a polymer bead in vitro before THBS1 treatment (Day 0; K) shows absence of sprouts. Arrows indicate representative sprouts on Day 2 of treatment with no THBS1 (L) and 0.1 nM THBS1 (M). Images were quantified for both the number of sprouts (sprouts/bead; G,H) and sprout length in μm (H,J). For (A,D,G–J), data are expressed as mean + SEM, n = 3–5 samples/treatment. Within each panel, groups with no common letters are different by ANOVA and Duncans post hoc test, p < 0.05.