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Supplemental Figure S1

Supplemental Figure S1. A. MDA-MB-231 BCL-2. MCF-7 BCL-2. MDA-MB-231. MCF-7. Bcl-2. Bcl-2. Actin. Actin. B. Supplemental Figure S2. **. *. *. *. *. Control. - - - - - - -. Cisplatin 30 M.

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Supplemental Figure S1

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

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

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

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

  5. Supplemental Figure S5 C A B

  6. Supplementary Figure Legends 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 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. 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 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 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.

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