Fig. 4

Sema3F is an inhibitor of angiogenesis in Matrigel® sprouting assays. (A-C) In cardiac endothelial cells (HCECs) Sema3F abolished the pro-angiogenic effect of VEGF. HCECs were cultured with the indicated amounts of Sema3F or VEGF alone or in combination over 16 to 18 h. HCECs were then assayed on Matrigel® for further 2.5 h. (A) Representative micrographs of stimulated HCECs are shown. (B&C) Sema3F prevented VEGF-induced cumulative sprout length (B) and branching of HCECs (C). Data from 4 independent experiments were shown (n, number of biological replicates). The results were analyzed using the one-way ANOVA and Bonferroni's multiple comparisons test: *p < 0.05. Figs. Dsingle bondF show the effect of Sema3F knockdown on angiogenesis of HCECs. The siRNA-based Sema3F knockdown resulted in a potentiated pro-angiogenic effect of VEGF. HCECs transfected with Sema3F-specific siRNA (siSema3F) or control siRNA (ctrl-siRNA) were treated with 100 ng/ml VEGF or EBM with 2 % FCS for 18 h before being assayed on Matrigel®. (D) Representative micrographs are shown. Sema3F knockdown led to a significant increase in cumulative sprout length (E) and number of branch points (F). (E&F) Data from 4 independent experiments were shown (n, number of biological replicates). All box plots extend from the 25th to the 75th percentile with the median marked. Whiskers extend from the 10th to the 90th percentile. The results were analyzed using one-way ANOVA followed by Bonferroni's multiple comparison test: *p < 0.05.