Attenuation of angiogenesis in vivo by 12-LOX inhibitors.

Matrigel implantation assay for angiogenesis was performed as described in “Materials and methods” and repeated at least 3 times with similar results. (A) Matrigel implantation angiogenesis assay. Top panel: left, bFGF (5 μg/mL Matrigel) in situ; right, bFGF (5 μg/mL Matrigel) resected Matrigel. Middle panel: left, bFGF (5 μg/mL Matrigel) and BHPP (0.8 mg/mL Matrigel) in situ; right, bFGF (5 μg/mL Matrigel) and BHPP (0.8 mg/mL Matrigel) resected implant. Bottom panel: left, Matrigel alone in situ; right, resected Matrigel blank control. (B) Hemoglobin levels in resected implants. The hemoglobin was measured by Drabkin's reagent. Columns, hemoglobin levels normalized with protein concentrations. Bars, SE from quadruplicate samples. (C) 12(S)-HETE levels in resected implants. Lipids were extracted from the resected implants, and 12(S)-HETE levels were measured as described in “Materials and methods.” Columns, average 12(S)-HETE levels normalized with protein concentrations. Bars, SE (n = 4 for Matrigel control; n = 3 for bFGF; n = 5 for bFGF plus BHPP).