Journal: PLoS ONE

Article Title: The Marine-Derived Oligosaccharide Sulfate (MdOS), a Novel Multiple Tyrosine Kinase Inhibitor, Combats Tumor Angiogenesis both In Vitro and In Vivo

doi: 10.1371/journal.pone.0003774

Figure Lengend Snippet: A–B, MdOS blocks HER-2 phosphorylation and its downstream signal transduction in SK-OV-3 cells (A) and NIH-3T3/neu cells (B). C–D, Inhibition of EGF-stimulated EGFR phosphorylation and signal transduction by MdOS in A431 (C) and MDA-MB-468 (D) cells. Serum-starved cells were incubated with indicated concentrations of MdOS for 6 h at 37°C, EGF (50 ng/ml) was added to the cultures during the last 15-min treatment. E–F, Inhibition of VEGF-stimulated VEGFR2 phosphorylation and signal transduction by MdOS. HMEC (E) and NIH-3T3/flk-1 (F) cells were starved, then incubated with indicated concentrations of MdOS for 6 h at 37°C, VEGF (50 ng/ml) was added to the cultures during the last 15-min treatment. Protein samples were separated by SDS-PAGE and probed using the indicated antibodies. Representative data are shown.

Article Snippet: Western blot analyses were subsequently performed as previous described , with the antibodies against phospho-EGFR (Tyr1068) (#2234), phospho-HER-2 (Tyr1248) (#2247), phospho-VEGFR2 (Tyr996) (#2474), VEGFR2 (#2472), phospho-ERK1/2 (#9101), ERK1/2 (#9102), phosphor-AKT (Ser473) (#9271) and AKT (#9272) (Cell Signaling Technology, Beverly, MA, USA), EGFR (sc-03), HER-2(sc-284) and actin (sc-8432) (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Transduction, Inhibition, Incubation, SDS Page

Journal: PLoS ONE

Article Title: The Marine-Derived Oligosaccharide Sulfate (MdOS), a Novel Multiple Tyrosine Kinase Inhibitor, Combats Tumor Angiogenesis both In Vitro and In Vivo

doi: 10.1371/journal.pone.0003774

Figure Lengend Snippet: VEGFR2 was injected at the concentrations of 2.56, 0.64, 0.32, 0.16 and 0.08 µM (from top to bottom); HER-2 was injected at the concentrations of 0.53, 0.26, 0.13, 0.07 and 0.03 µM (from top to bottom); EGFR was injected at the concentrations of 6, 3, 1.5, 0.75, 0.38, 0.19 and 0.09 µM (from top to bottom); 6×his-tag was injected at the concentrations of 100, 10, 1, 0.1, 0.01 µM. Sensorgram responses at equilibrium were plotted against each concentration of compounds and the equilibrium dissociation constant (K D ) of the binding system was calculated using BIAeval software 3.1.

Article Snippet: Western blot analyses were subsequently performed as previous described , with the antibodies against phospho-EGFR (Tyr1068) (#2234), phospho-HER-2 (Tyr1248) (#2247), phospho-VEGFR2 (Tyr996) (#2474), VEGFR2 (#2472), phospho-ERK1/2 (#9101), ERK1/2 (#9102), phosphor-AKT (Ser473) (#9271) and AKT (#9272) (Cell Signaling Technology, Beverly, MA, USA), EGFR (sc-03), HER-2(sc-284) and actin (sc-8432) (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Injection, Concentration Assay, Binding Assay, Software

Journal: PLoS ONE

Article Title: The Marine-Derived Oligosaccharide Sulfate (MdOS), a Novel Multiple Tyrosine Kinase Inhibitor, Combats Tumor Angiogenesis both In Vitro and In Vivo

doi: 10.1371/journal.pone.0003774

Figure Lengend Snippet: VEGFR2 (A) and HER-2 (B) and EGFR (C) kinase assays were performed as described in in the presence of varying concentrations of ATP. Initial reaction velocity was expressed as the phosphorylation of poly(Glu, Tyr) 4∶1 substrate. All x, y data sets were multiplied by 100 for purposes of graphical presentation.

Article Snippet: Western blot analyses were subsequently performed as previous described , with the antibodies against phospho-EGFR (Tyr1068) (#2234), phospho-HER-2 (Tyr1248) (#2247), phospho-VEGFR2 (Tyr996) (#2474), VEGFR2 (#2472), phospho-ERK1/2 (#9101), ERK1/2 (#9102), phosphor-AKT (Ser473) (#9271) and AKT (#9272) (Cell Signaling Technology, Beverly, MA, USA), EGFR (sc-03), HER-2(sc-284) and actin (sc-8432) (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: