Figure 6: ROBO4ΔCD does not affect Slit2-mediated AKT activation and sprouting.
From: The Robo4 cytoplasmic domain is dispensable for vascular permeability and neovascularization
(a) Schematic of possible ROBO4 and Robo4ΔCD effects on Slit2-ROBO1/2 signalling. (b) Western blot of phospho-AKT S473 (p-AKT) (left) and quantification (right) in HUVECs with Ctrl or ROBO4 siRNA transfection and 6 nM Slit2 stimulation. #: Slit2 significantly induces p-AKT at 30 min. N=3 experiments. Error bars: s.e.m. NS, not significant, Student's t-test. (c) Left panel: western blot analysis of ROBO4 expression and p-AKT in HUVECs with indicated siRNA transfection and adenovirus infection after 6 nM Slit2 stimulation. Right panel: western blot quantifications of p-AKT/tot-AKT (n=4). #: Slit2 significantly induces p-AKT at 30 min. Error bars: s.e.m. NS, not significant, Student's t-test. (d) HUVEC sprouting in 3D fibrin gels (left) and the corresponding quantifications (right, n=3). Cells were treated with siRNAs and virus as indicated and then stimulated with 6 nM Slit2 for 96 h. #: Slit2 significantly induces HUVEC sprouting. Error bars: s.e.m. NS, not significant, Mann–Whitney U test. Scale bar, 200 μm. (e) Western blot analysis of p-AKT in HUVECs with indicated siRNA transfection and adenovirus infection after 6 nM Slit2 stimulation. N=3 experiments. #: Slit2 significantly induces p-AKT at 30 min. Error bars: s.e.m. * P<0.05 when compared with Ctrl siRNA+adGFP group, Student's t-test.
