Effect of FAE in regulating Bax translocation.

<p>(<b>A</b>) MCF-7 cells expressing Bax EGFP and Mito DsRed were seeded in 96 well glass bottom plate (BD, USA) with low density and after 48 h, treated with FAE at 100 µg/ml. The image was taken using BD Pathway Bioimager 435 (Becton Dickinson, USA) at 3, 18 and 27 h by setting Montage (2×2) and specific Macro using AttoVision™ software. The green granular aggregate indicates the translocation of Bax to mitochondria, as indicated by the arrows. The representative images collected at indicated time points were used for analysing the percentage positive cells with Bax EGFP onto mitochondria compared to total in the field. (<b>B</b>) Graph showing the percentage of cells undergoing Bax translocation into mitochondria upon FAE treatment. (<b>C</b>) The MCF-Bax-DS Red cells were treated as indicated above. Bax-EGFP accumulation in mitochondria is indicated in high magnification images with arrows. A magnified merged image of the treated cells is also shown. (<b>D</b>) MCF-7 cells were transfected with Control (scr) siRNA or Bax siRNA. After 48 h of transfection, whole cell extract was prepared and immunoblotted for Bax and Actin. The same cells were also stained with Hoechst 33342 after 48 h of FAE treatment to visualize chromatin condensation (left panel). The graph is the quantitative representation of the percentage of cells with condensed chromatin in scrambled-transfected and Bax-transfected cells after FAE treatment.