PMC

(A) The effects of gedunin (1–20 μM) on the expression profile of apoptotic protein expression profiles such as PARP, cleaved caspase 3, Bcl‐xL, and BAD in U87 and U251. The cells were treated with gedunin for 72 h. These figures are representative images of three biological replicates. (B) The effects of gedunin (1–20 μM) on apoptotic protein expression profiles such as PARP, cleaved caspase 3, Bcl‐xL, BAD in U87 EGFR vIII, U251 EGFR vIII, and U251 EGFR. The cells were treated with gedunin for 72 h. These figures are representative images of three biological replicates. (C) Caspase 3 inhibitor Ac‐DEVD‐CHO attenuates the effects of gedunin (0–20 μM) on the expression of cleavage of PARP in U251 MG and U87 MG cells. Caspase 3 inhibitor Ac‐DEVD‐CHO abrogates the decrease in cell viability induced by gedunin (0–20 μM) in U251 MG and U87 MG cells. These figures are representative images of three biological replicates. These results are not statistically significant. (D) Effects of gedunin on cell apoptosis in U251 MG and U87 MG cells. Gedunin (5–20 μM) induced significant (p < .05) apoptotic events using the BD Accuri C6 plus. These figures are representative images of three biological replicates.

Apoptotic NTERA-2 cells after exposure to gedunin evaluated by TUNEL assay. (a) Untreated control, (b) cells treated with 2.5 μg/mL of gedunin, (c) cells treated with 5 μg/mL of gedunin, and (d) cells treated with 10 μg/mL of gedunin.

Morphological changes observed under florescence microscope in gedunin treated NTERA-2 cells after staining with AO/EB. (a) Untreated control, (b) cells treated with 2.5 μg/mL of gedunin, (c) cells treated with 5 μg/mL of gedunin, and (d) cells treated with 10 μg/mL of gedunin.

A. C. E. and G. Cells were treated with 5-50μM gedunin (GD). Cell viability was assessed using MTS analysis after treatment with different doses of gedunin for 24h. B. D. F. and H. Control and 25μM gedunin treated cells (their IC₅₀ concentration) were visualized under light microscope at 10X magnification. I. Western blot analysis of total and phosphorylated forms of AKT, PI3K, mTOR, and p70s6kinase in pancreatic cancer cell lysates treated with 25μM gedunin. Data are expressed as the mean ± SEM (*p<0.05) of three separate experiments.