Angiogenesis Inhibition Assay Protocol

Ethanol, fibrinogen, aprotinin, thrombin and Pyrrolidine dithiocarbamate (PDTC) were purchased from Sigma Chemical Co. (St. Louis, MO). Cyanidin-3-glucoside (C3G) was prepared as previously described [38 (link)]. Anti-MCP1 and anti-VEGF antibodies were obtained from Abcam (Cambridge, MA). Anti-NF-κB p65, IκBα, p-IκBα and anti-LMNB1 antibodies were purchased from Protein Tech Group (Chicago, IL, USA). Anti-β-actin was obtained from Cell signaling Technology (Danvers, MA). Anti-CD31 antibody was obtained from BD Pharmingen (San Diego, CA). Reactive oxygen species detection reagents were obtained from Invitrogen Molecular Probes (Eugene, OR). MTT assay kit was purchased from Roche Molecular Biochemicals (Indianapolis, IN). Matrigel Invasion Chambers were purchased from BD Biosciences (Bedford, MA). Cytodex 3 beads were purchased from Amersham Pharmacia Biotech (Piscataway, NJ).