Figure 4.

IDET induces apoptosis in breast cancer cells. (A) MDA-MB-231 and MCF-7 cells were treated with 5, 10 and 25 µM IDET. After 24 hrs, cells were stained with AO/PI, washed and examined under fluorescence microscope. (B) Untreated and IDET treated MDA-MB-231 cells were stained with PI, and flow cytometry was used to analyze the cells at different phases of cell cycle. (C) IDET induces PS externalization in breast cancer cells. MDA-MB-231 cells were exposed to 10 µM IDET for 24 hrs, stained with Alexafluor 488 conjugated annexin V antibody and analyzed by flow cytometry. (D) DNA was extracted from control and treated cells and electrophoresed on 1.5% agarose gel containing ethidium bromide. (E) The whole cell extract obtained from untreated and treated MDA-MB-231 cells was used to examine the expression pattern of cell survival, invasive, cleaved caspase, and PARP proteins. The corresponding GAPDH and β-Actin was used for the normalization of the data. The fold reduction in the experimental group as compared to the control group is indicated below the blot. (F) The RNA was extracted from the untreated and treated cells, reverse transcribed, amplified by PCR, electrophoresed on the agarose gel and stained with ethidium bromide. The blots were derived from the same gel and the data was normalized using GAPDH as an internal control. IDET, isodeoxyelephantopin.