Matrigel plug angiogenesis assay

Precooled growth factor-reduced Matrigel was mixed with 1 μg/mL and 10 μg/mL AMEs and 1 μg/mL AMEs combined with 10 μM PDTC at a ratio of 1:1.^17,30 To generate Matrigel plugs, 300 µL of this mixture was injected subcutaneously into the dorsal region of nude mice. After 2 weeks, a Drabkin’s reagent kit (Sigma-Aldrich) was used to evaluate the hemoglobin content. ELISA was used to analyze the expression of growth factors, and protocols described by the provider for PDGF-BB and VEGF (Cloud-Clone) were used for this assay. WB was performed as described previously to analyze the expression of NFκb-associative proteins, and the images were assessed via ImageJ software (National Institutes of Health).