Figure 4
From: MEG8 regulates Tissue Factor Pathway Inhibitor 2 (TFPI2) expression in the endothelium
TFPI2 silencing rescues impaired sprouting after loss of MEG8. MEG8 and TFPI were knocked down by GapmeR and siRNA, respectively. (A) EC spheroids were embedded in collagen gels 24 h after transfection and stimulated with VEGF. Spheroid were fixed after 24 h VEGF stimulation. Cumulative sprout length was determined by measuring the distance from the base of the spheroid to the tip cell. Discontinuous sprouts were excluded. For each independent experiment 7–15 spheroids were scored. 10 independent experiments were performed. Scale bar indicates 100 µm. Multiple groups were analysed using one-way ANOVA. (B) Proliferation was measured by EdU incorporation between 24–48 h after transfection. The percentage of EdU-positive cells is shown. Multiple groups were analysed using one-way ANOVA (C) In addition, cell numbers were counted over time. HUVECs were seeded at equal densities and counted 24-, 48- and 72 h after transfection. Data are presented as mean ± SEM. Multiple groups were analysed using one-way ANOVA. Significance was indicated as: *p < 0.05, **p < 0.01, ***p < 0.001.
