AE treatment induces autophagy, and inhibits angiogenesis and Hif-1α expression in xenografted tumors.

<p>A. Photograph showing increased immunoexpression of beclin1 in xenografted tumors after being treated with AE. The histogram shows the percentage of beclin1 immunopositive cells compared to total cells in control and AE treated groups. B. Representative photograph of a Western blot showing increased expression of beclin1 in AE treated group. C. Representative photograph of a Western blot showing increased expression of LC3B-II in AE treated group. β-actin was used as a control for each blot. Mean <u>+</u> S.E.M. values of densitometric ratio of respective protein (beclin1 and LC3B-II) and β-actin are shown on the bottom of each gel. D. AE treatment decreased immunohistochemical expression of CD31 positive cells in mouse xenograft tumors. Histogram showing decreased microvessel density in xenografts of AE-treated mice. E. Photomicrograph showing reduced expression of HIF-1α immunostaining in xenograft tumors after AE treatment. The histogram shows the percentage of Hif-1α immunopositive cells compared to total cells. F. Representative photograph of a Western blot for Hif-1α is shown on the top. β-actin was used as a control for each blot. Mean <u>+</u> S.E.M. values of densitometric ratio of Hif-1α and β-actin are shown on the bottom of the gel. After immunodetection, the density of Hif-1α positive bands was measured and normalized with β-actin values. Tissue sections were photographed at 400X magnification. The values are means <u>+</u> S.E.M. of 5 different mice. *, p<0.05, as compared with control group. Bar=50 µm.