Analysis of apoptosis in MDA-MB 231 and primary breast adenocarcinoma cells.

<p>(A) Cells treated with vehicle and CDRI-85/287 were analyzed by flow cytometry of annexin-V/PI stained cells after 48h culture. AV⁺/PI⁻ - intact cells; AV⁻/PI⁺ -nonviable/necrotic cells; AV⁺/PI⁻ and AV⁺/PI⁺ - apoptotic cells. Representative images are shown in the upper panel and the percentage of cell fraction with SEM based on three independent experiments is shown in the lower panel. p values are a-p<0.001, b-p<0.01, c-p<0.05 and d-p>0.05 vs. control. (B) Western blot analysis to see the expression of pro- and anti- apoptotic proteins. MDA-MB 231 and primary breast cancer cells were treated with the indicated concentrations of compound for 48h, and 30 µg whole cell lysate in each lane was probed for the expression of different proteins using specific antibodies. β-actin was used as a control to correct for loading. Representative blots are shown in the upper panel and densitometric quantitation of protein expression levels are shown as fold changes in the lower panel. Results are expressed as mean ± SEM, n = 3. p values are a-p<0.001, b<0.01, c-p<0.05 and d-p>0.05 vs. control.