Figures & data
Figure 1 Synthesis and 1H-NMR spectra of CS-ES2-AF. (A) Synthetic route of CS-ES2-AF. 1H-NMR spectra of (B) CS, (C) CS-TBA, and (D) CS-ES2-AF.
Abbreviations: CS, chondroitin sulfate; 1H-NMR, 1H-nuclear magnetic resonance spectroscopy.
![Figure 1 Synthesis and 1H-NMR spectra of CS-ES2-AF. (A) Synthetic route of CS-ES2-AF. 1H-NMR spectra of (B) CS, (C) CS-TBA, and (D) CS-ES2-AF.Abbreviations: CS, chondroitin sulfate; 1H-NMR, 1H-nuclear magnetic resonance spectroscopy.](/cms/asset/6c0106a6-16d2-4069-b457-89e971f7b0be/dijn_a_195934_f0001_c.jpg)
Figure 2 Characterization of CS-ES2-AF nanoparticles. (A) TEM image of CS-ES2-AF and ES2-AF nanoparticles. (B) Size distribution of CS-ES2-AF and ES2-AF nanoparticles.
Abbreviations: CS, chondroitin sulfate; TEM, transmission electron microscopy.
![Figure 2 Characterization of CS-ES2-AF nanoparticles. (A) TEM image of CS-ES2-AF and ES2-AF nanoparticles. (B) Size distribution of CS-ES2-AF and ES2-AF nanoparticles.Abbreviations: CS, chondroitin sulfate; TEM, transmission electron microscopy.](/cms/asset/3a71c259-b2d8-41a2-89b6-67d8a1cac843/dijn_a_195934_f0002_b.jpg)
Figure 3 (A) Inhibitory effects of peptide ES2-AF and conjugate CS-ES2-AF on the proliferation of EA.hy926 endothelial cells after incubation for 48 hours. (B) Comparison of the effects of ES2-AF, the mixture of CS&ES2-AF, and CS-ES2-AF conjugation based on in vitro ELISA assay. Data represent mean ± SD (n=5). **P<0.01 is considered significantly different compared with the ES2-AF group.
Abbreviation: CS, chondroitin sulfate.
![Figure 3 (A) Inhibitory effects of peptide ES2-AF and conjugate CS-ES2-AF on the proliferation of EA.hy926 endothelial cells after incubation for 48 hours. (B) Comparison of the effects of ES2-AF, the mixture of CS&ES2-AF, and CS-ES2-AF conjugation based on in vitro ELISA assay. Data represent mean ± SD (n=5). **P<0.01 is considered significantly different compared with the ES2-AF group.Abbreviation: CS, chondroitin sulfate.](/cms/asset/a84fa3a9-e4f8-48c2-8fe1-800093f40f6e/dijn_a_195934_f0003_c.jpg)
Figure 4 Effects of different samples on the inhibition of cell migration, tube formation, and CAM: (i) represents the ES2-AF sample; (ii) represents the CS-ES2-AF sample; and (iii) represents the CS&ES2-AF sample. (A) Cell migration assay. (B) Tube formation assay. (C) CAM assay. Comparison of different samples for (D) cell migration and (E) tube formation. (F) Comparison of the inhibition of ES2-AF and its derivatives and the mixture on CAM angiogenesis. Data represent mean ± SD (n=5). #P<0.05 is considered significantly different compared with the control group. ##P<0.01 is considered significantly different compared with the control group. *P<0.05 is considered significantly different compared with the ES2-AF group. **P<0.01 is considered significantly different compared with the ES2-AF group.
![Figure 4 Effects of different samples on the inhibition of cell migration, tube formation, and CAM: (i) represents the ES2-AF sample; (ii) represents the CS-ES2-AF sample; and (iii) represents the CS&ES2-AF sample. (A) Cell migration assay. (B) Tube formation assay. (C) CAM assay. Comparison of different samples for (D) cell migration and (E) tube formation. (F) Comparison of the inhibition of ES2-AF and its derivatives and the mixture on CAM angiogenesis. Data represent mean ± SD (n=5). #P<0.05 is considered significantly different compared with the control group. ##P<0.01 is considered significantly different compared with the control group. *P<0.05 is considered significantly different compared with the ES2-AF group. **P<0.01 is considered significantly different compared with the ES2-AF group.](/cms/asset/369d67e9-830e-4e23-8ef8-f2a8c1012159/dijn_a_195934_f0004_c.jpg)
![Figure 4 Effects of different samples on the inhibition of cell migration, tube formation, and CAM: (i) represents the ES2-AF sample; (ii) represents the CS-ES2-AF sample; and (iii) represents the CS&ES2-AF sample. (A) Cell migration assay. (B) Tube formation assay. (C) CAM assay. Comparison of different samples for (D) cell migration and (E) tube formation. (F) Comparison of the inhibition of ES2-AF and its derivatives and the mixture on CAM angiogenesis. Data represent mean ± SD (n=5). #P<0.05 is considered significantly different compared with the control group. ##P<0.01 is considered significantly different compared with the control group. *P<0.05 is considered significantly different compared with the ES2-AF group. **P<0.01 is considered significantly different compared with the ES2-AF group.](/cms/asset/18d10c2d-9f46-401d-89f8-e9c4589ed32d/dijn_a_195934_f0004a_c.jpg)
Figure 5 Binding ability of CS-ES2-AF to the CD44 receptor in vitro. (A) pH scouting of CD44 in acetate buffer solution. (B) SPR sensorgram of different concentrations of CS-ES2-AF over a chip with immobilized CD44. (C) SPR sensorgram of different concentrations of ES2-AF over a chip with immobilized CD44.
Abbreviations: CS, chondroitin sulfate; SPR, surface plasmon resonance.
![Figure 5 Binding ability of CS-ES2-AF to the CD44 receptor in vitro. (A) pH scouting of CD44 in acetate buffer solution. (B) SPR sensorgram of different concentrations of CS-ES2-AF over a chip with immobilized CD44. (C) SPR sensorgram of different concentrations of ES2-AF over a chip with immobilized CD44.Abbreviations: CS, chondroitin sulfate; SPR, surface plasmon resonance.](/cms/asset/0bd7a612-807c-4d26-a9d9-3391c94c668f/dijn_a_195934_f0005_c.jpg)
Figure 6 In vivo tumor-targeting ability of CS-ES2-AF in nude mice bearing B16 tumor xenografts, monitored by the Optix system. Tumor is indicated using red circle.
Abbreviation: CS, chondroitin sulfate.
![Figure 6 In vivo tumor-targeting ability of CS-ES2-AF in nude mice bearing B16 tumor xenografts, monitored by the Optix system. Tumor is indicated using red circle.Abbreviation: CS, chondroitin sulfate.](/cms/asset/2a992b9f-0054-468c-bc97-ac8f357d9974/dijn_a_195934_f0006_c.jpg)
Figure 7 Pharmacokinetic time curves of ES2-AF and CS-ES2-AF in plasma after single intravenous administration. The results are shown as means ± SD (n=5).
Abbreviation: CS, chondroitin sulfate.
![Figure 7 Pharmacokinetic time curves of ES2-AF and CS-ES2-AF in plasma after single intravenous administration. The results are shown as means ± SD (n=5).Abbreviation: CS, chondroitin sulfate.](/cms/asset/292287a3-9e1d-4655-b1da-55361393a73e/dijn_a_195934_f0007_c.jpg)
Table 1 Pharmacokinetic parameters of plasma concentration of ES2-AF and CS-ES2-AF in mice