Fig. 3

Effect of Au(FeCo) and (FeCo) nanoparticles on VEGF165-induced proliferation of HUVECs at various concentrations (0.1–10 µg/mL), observed by [³H]thymidine incorporation (cell proliferation) assay, presented as fold of stimulation. a VEGF165-induced proliferation of HUVECs in the presence of Au(FeCo). Au(FeCo) nanoparticles were pre-incubated with and without VEGF (10 ng/mL) for 30 min at 4°C, added to HUVEC starved with (0.1% serum) for 24 h in 24-well plates, and incubated for 24 h. After 24 h of incubation with nanoparticles, 1 μCi [³H]thymidine was added into each well. Four hours later, cells were washed with cold PBS, fixed with 100% cold methanol, and the lysate collected with 0.1(N) NaOH for measurement of radioactivity. Experiments were repeated in triplicate. b VEGF165-induced proliferation of HUVECs in the presence of (FeCo). Experiments were performed similarly as described above. c Western blot analysis of VEGFR2 upon nanoshell treatment. Effect of Au(FeCo) and (FeCo) on receptor phosphorylation. Western blot analysis of phopho-KDR (p-VEGFR2) and total KDR (VEGFR2) from (1) control untreated HUVECs, (2) A, serum-starved HUVECs were stimulated for 5 min withVEGF (10 ng/mL) as a positive control, (3–5) HUVECs treated with Fe–Co–Au at various concentrations [3 = 100 ng, 4 = 1 µg, and 5 = 10 µg of Fe–Co–Au], and (6–8) HUVECs treated with Fe–Co at various concentrations [6 = 100 ng, 7 = 1 µg, and 8 = 10 µg of Fe–Co]