Fig 2: Elevated expression of GPC1 is associated with clinicopathological features of CRC.(A–J). GPC1 and GPC2 mRNA expression analysis in TCGA CRC samples (453 samples) by age, stage, T, N and M using Kolmogorov–Smirnov test. (K). Immunohistochemical staining for GPC1 in CRC tissues and normal tissues.

Fig 3: HDMEC express the receptors for VEGF and HGF, which are secreted by KORS. (A) Growth factors secreted by KORS analyzed by protein array from their conditioned medium (KORSCM) and compared to the basal cell culture medium (ECM) without cells. Green frames: positive control spots; red frames: negative control spots; black frames: spots corresponding to the indicated target. (B) The production of HGF and VEGF in the KORS were compared to that of the HHFDPCs by Western immunoblotting. (C) Growth factors and receptors of HDMECs analyzed by protein array. Green frames: positive control spots; red frames: negative control spots; black frames: spots corresponding to the indicated target. (D) The direct interaction between GPC1 with VEGFR2 or c-Met was analyzed by co-immunoprecipitation. A whole cell protein extract (25 µg) from the HDMECs was precipitated using anti-GPC1 antibody. Anti-c-Met or anti-VEGFR2 antibody was used to reveal the membrane. A control experiment without antibody was performed. (E) A reverse immunoprecipitation assay, corresponding to the co-immunoprecipitation shown in Figure 8D, was conducted from HDMEC whole cell protein extracts using anti-VEGFR2 or c-Met antibody. Then, the isolated immunocomplexes were immunoblotted using anti-GPC1 antibody.

Fig 4: Schematic representation of interactions between GPC1, c-Met and VEGFR2 and their ligands at the HDMEC membrane. (A) The downstream effect of the binding of the ligands secreted by KORS to their specific receptors expressed on HDMECs and the interaction between the receptors are illustrated. The effects of VEGF and HGF on HDMEC pseudotube formation are presented. (B) Effects of GPC1 cleavage by PLC on HDMEC pseudotube formation and proliferation are illustrated.

Fig 5: Glypican-1 knockout mice have inhibited eNOS phosphorylation in both chronic and increased flow conditions. Descending aorta from WT and Gpc1-/- mice were en face immunostained for peNOS and PECAM-1. (A) Mean normalized peNOS expression under chronic flow is shown for WT (n = 10) and Gpc1-/- mice (n = 10). *P < 0.05 using two-tailed student’s t-test. (B) Mean normalized PECAM-1 expression under chronic flow is shown for WT (n = 6) and Gpc1-/- (n = 5) mice. *P < 0.05 using two-tailed student’s t-test. (C) Mean normalized peNOS expression under increased flow induced by dobutamine for 30 min is shown for WT (n = 9 and 8 for saline and dobutamine groups, respectively) and Gpc1-/- mice (n = 8 for both saline and dobutamine groups). *P < 0.05 using one-way ANOVA followed by Sidak’s multiple comparisons test.