1. Supplemental Figure S1 A B MDA-MB-231 MCF-7 BCL-2MDA-MB-231 BCL-2 MCF-7 Bcl-2 Actin Bcl-2 Actin

2. - - - - - - - Control Cisplatin 30  M TEMPO 2 mM Trolox 200  M U-74389G 5  M Tiron 10 mM - + + + + + + - - + - - - - NAC 10 mM - - - + - - - - - - - - + - - - - - - - + - - - - + - - Supplemental Figure S2 ** ** **

3. IP: Bcl-2 IB: 4-HNE IP: Bcl-2 IB: 4-HNE IP: Bcl-2 IB: Bcl-2 IP: Bcl-2 IB: Bcl-2 MCF-7 BCL-2 MCF-7 - + Cisplatin 30  M - + Cisplatin 30  M Supplemental Figure S3

4. Bak Actin IP: Bak Ab-2 IB: Bak Input IB: Bak MCF-7 BCL-2 Trolox 200  M U-74389G 5  M - - + - - - - + - + + + - - - - Cisplatin 30  M Control Supplemental Figure S4

5. Supplemental Figure S5 AB C

6. Supplementary Figure S1. A, Immunoblot analysis of Bcl-2 expression in parental and Bcl-2-transfected MCF-7 and MDA- MB-231 cells. B, MCF-7 and MCF-7 Bcl-2 cells were treated with paclitaxel (20 nM), cisplatin (30  M) and HA14-1 (20  M) for 48 h and apoptosis was evaluated by M30 Apoptosense assay. Supplementary Figure S5. MCF-7 Bcl-2 cells were transfected with Noxa siRNA or scramble siRNA. Cells were treated with cisplatin (30  M) for 48 h. A, apoptosis was evaluated by M30 Apoptosense ELISA. B, MMP loss was evaluated by flow cytometry using MitoTracker Red CMXRos. C, caspase-9 activation was determined by fluorometric caspase assay. Supplementary Figure Legends Supplementary Figure S4. MCF-7 Bcl-2 cells were preincubated with lipid peroxidation inhibitors (Trolox, U-74389G) for 1 h and then treated with cisplatin for 36 h. Activation of Bak was assessed by immunoprecipitation using active conformation- specific anti-Bak (Ab-2) antibody followed by immunoblot analysis. 5% of the input for immunoprecipitation was also subjected to immunoblot analysis. Actin was used as a loading control. Supplementary Figure S3. MCF-7 and MCF-7 Bcl-2 cells were treated with cisplatin (30  M) for 12 h. (4-HNE)-histidine adduct formation was assessed by immunoprecipitation using anti-Bcl-2 (#2872) antibody followed by immunoblot analysis using anti-HNE monoclonal antibody. Immunoprecipitated proteins were also probed with anti-Bcl-2 antibody to test the efficiency of immunpoprecipitation experiments. Supplementary Figure S2. MCF-7 Bcl-2 cells with cisplatin (30  M) or cisplatin plus NAC (10 mM), TEMPO (2 mM), Tiron (10 mM), Trolox (200  M) or U-74389G (5  M) for 48 h. Apoptosis was detected by Annexin V staining. Columns, mean of three independent experiments; bars, SE. *, P < 0.05, **, P < 0.01.