Figure 6.

TanFe (transcellular activator of nuclear FOXF1 expression) increases angiogenesis in FOXF1 (Forkhead Box F1)-deficient endothelial cells in vitro. (A) Western blot shows that TanFe treatment increases FOXF1 protein concentrations but does not affect Vinculin in FOXF1-deficient endothelial Mouse Fetal Lung Mesenchyme-91 U (MFLM-91U) cells. Inhibition of FOXF1 was achieved by transient transfection with FOXF1-specific siRNA (siFOXF1). siRNA against a nontargeted RNA sequence was used as a control (siCtrl). (B) Quantification of FOXF1 protein concentrations was performed using densitometric analysis of Western blot images and normalized to Vinculin (n = 3). (C–E) In vitro angiogenesis assay shows that TanFe increases the formation of endothelial sprouts in endothelial Human umbilical vein endothelial cells (HUVECs) infected with shFOXF1 lentivirus. Lentiviral particles with shRNA against a nontargeted RNA sequence were used as a control (shCtrl). Scale bars, 100 μm. The complexity of the vascular network in Matrigel was quantitated by measurements of sprout length and counts of the sprout junctions (n = 4 for each group). *P < 0.05. n.s. = not significant.