I. Kausch, H. Jiang, B. Thode, C. Doehn, S. Krüger, D. Jocham

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Inhibition of bcl-2 Enhances the Efficacy of Chemotherapy in Renal Cell Carcinoma I. Kausch, H. Jiang, B. Thode, C. Doehn, S. Krüger, D. Jocham European Urology Volume 47, Issue 5, Pages 703-709 (May 2005) DOI: 10.1016/j.eururo.2004.11.013 Copyright © 2004 Elsevier B.V. Terms and Conditions

Fig. 1 bcl-2 expression in different cell lines. (a) RT-PCR analysis of bcl-2 and GAPDH mRNA expression in different cell lines. (Three RCC cell lines: SK-RC 35, 47, and 58). (b) Western blot analysis of bcl-2 and actin protein expression. One representative experiment is shown. European Urology 2005 47, 703-709DOI: (10.1016/j.eururo.2004.11.013) Copyright © 2004 Elsevier B.V. Terms and Conditions

Fig. 2 bcl-2 expression after antisense and control treatment. (a) RT-PCR and (b) Western blot analysis of bcl-2 expression in SK-RC 58 cells was performed after transfection of SK-RC 58 cells with bcl-2 antisense (AS), control (MM) oligonucleotides or Tfx control only (Tfx) at a concentration of 500nM compared to untreated cells (H2O). Experiments were performed in triplicate. One representative experiment is shown. European Urology 2005 47, 703-709DOI: (10.1016/j.eururo.2004.11.013) Copyright © 2004 Elsevier B.V. Terms and Conditions

Fig. 3 MTT test after bcl-2 antisense mono-treatment. Incubation of SK-RC 58 cells with different concentrations of antisense (AS) or control (MM) oligonucleotides as well as Tfx only showed that no direct effect on cell viability was observed. European Urology 2005 47, 703-709DOI: (10.1016/j.eururo.2004.11.013) Copyright © 2004 Elsevier B.V. Terms and Conditions

Fig. 4 MTT test after combined bcl-2 antisense and cisplatin treatment. SK-RC 58 cells were transfected with 500nM antisense (AS), control ON (MM) or Tfx only. After 24hours Cisplatin was added at different concentrations. MTT assay demonstrates significantly increased reduction of cell viability after combined antisense and cisplatin treatment. Experiments were reproduced 3 times. Figure shows one representative result. European Urology 2005 47, 703-709DOI: (10.1016/j.eururo.2004.11.013) Copyright © 2004 Elsevier B.V. Terms and Conditions

Fig. 5 Apoptosis after combination therapy. Apoptotic cells identified by immunohistochemistry with the M30 antibody. Pictures show representative staining with high positivity after combined bcl-2 antisense (AS) ON (500nM) and cisplatin (1μg/ml) treatment and low positivity after combined treatment with control ON (MM). European Urology 2005 47, 703-709DOI: (10.1016/j.eururo.2004.11.013) Copyright © 2004 Elsevier B.V. Terms and Conditions

Fig. 6 Dependence of effectivity on bcl-2 expression. SK-RC 47 cells were transfected with 500nM antisense (AS), control ON (MM) or Tfx only. After 24hours Cisplatin was added at different concentrations. MTT assay shows only minor reduction of cell viability after combined antisense and cisplatin treatment. Experiments were reproduced 3 times. Figure shows one representative result. European Urology 2005 47, 703-709DOI: (10.1016/j.eururo.2004.11.013) Copyright © 2004 Elsevier B.V. Terms and Conditions