Fig. 6: VEGF maintains LSECs fenestrae via inhibiting autophagic degradation of Cav-1.
From: Autophagic degradation of caveolin-1 promotes liver sinusoidal endothelial cells defenestration
Primary rats LSECs, isolated from normal rats and cultured for 3 days in vitro, were treated with VEGF at the dose of 10 ng/ml from 12 h till 72 h. a Protein levels of VEGFR2, p-PI3K (Tyr458), PI3K, p-AKT (Ser473), AKT, p-MTOR (Ser2448), and MTOR analyzed by western blot. The relative protein expression is quantified in the graph, down. *P < 0.05 versus VEGFR2/β-actin of the concurrent control group; #P < 0.05 versus p-PI3K/PI3K of the concurrent control group; &P < 0.05 versus p-AKT/AKT of the concurrent control group; $P < 0.05 versus p-MTOR/MTOR of the concurrent control group. b Red or yellow represents autolysosomes or autophagosomes, respectively, visualized by confocal microscopy. Quantification of autophagic flux (%) in 100 cells was analyzed. *P < 0.05 versus the autolysosomes of the concurrent control group; #P < 0.05 versus the autophagosomes of the concurrent control group. c Protein levels of LC3 II/I, Cav-1, p-eNOS (Ser1177), eNOS, p-VASP (Ser157), and VASP in primary LSECs analyzed by western blot. The relative protein expression is quantified in the graph, down. *P < 0.05 versus LC3 II/I of the concurrent control group; #P < 0.05 versus Cav-1/GAPDH of the concurrent control group; &P < 0.05 versus p-eNOS/eNOS of the concurrent control group; $P < 0.05 versus p-VASP/VASP of the concurrent control group. d Magnification of SEM of LSECs in the CTR group and the VEGF group on Day 3, revealing the fenestrae structures (scale bar: 5 μm), and quantification of the total fenestral diameter, right. The black triangles indicate LSECs fenestrae structures. *P < 0.05 versus the concurrent control group. e The co-localization of LC3 (green) with Cav-1 (red) and F-actin (blue) in LSECs of the two groups (CTR and VEGF), shown by immunofluorescence. Scale bar: 10 μm
