Figure 5.

Ery5 inhibits angiogenesis and induces autophagy in vivo. (a) C57/BL6J mice (4- to 6-week old) were obtained from central animal facility of Indian Institute of Integrative Medicine, Jammu. Matrigel was embedded in mice as described in Materials and Methods. Animals were treated with Ery5 50 mg/kg, 100 mg/kg and sunitinib 2.5 mg/kg. Animals were killed on 10th day of experiment, matrigel plug was extracted and photographs were taken as described in Materials and Methods. (b) Hemoglobin content was measured from tissue samples as described in Materials and Methods. There was significant decrease in hemoglobin content in samples treated with Ery5, which represent its anti-angiogenic activity. Data are mean±S.D. of three similar experiments; statistical analysis was done by using the Bonferroni method and P-value<0.05 was considered to be significant with ***P<0.001. (c) Tissue samples from mice (matrigel) were homogenized in a homogenizer, followed by lysing in RIPA buffer. Protein estimation was done by the Bradford method, 60 μg protein was loaded in each well. Western blot for indicated antibodies was done as described in Materials and Methods. β-actin was used as an internal control