A B C Supplementary Figure S1. Trabectedin decreases viability of primary MPM cell cultures. A and B, dose-dependent impact of trabectedin on epithelioid (A) and non-epithelioid (B) MPM cell cultures established from surgical patient samples. C, Activity of trabectedin and cisplatin (IC50 values) grouped according to the histological origin (compare also Supplementary Table S1). Hoda et al

Supplementary Figure S2. Impact of trabectedin on MPM cell migration Supplementary Figure S2. Impact of trabectedin on MPM cell migration. Cells were exposed to the indicated trabectedin concentrations and cell migration was assessed by wound closure assays for the indicated time frames. Hoda et al

Supplementary Figure S3 Supplementary Figure S3. Activity of trabectedin combined with cisplatin against MPM cell lines. MPM cell lines were continuously exposed for 72 hours to the indicated doses of trabectedin and cisplatin. Changes in viability were measured by MTT assay (compare Figure 5) and data are shown as combination indices (CI). CI values <0.9, from 0.9 to 1.1, or >1.1 represent synergism, additive effects, and antagonism, respectively. Hoda et al

A * B Supplementary Figure S4. A, Expression of bcl-2 mRNA (data derived from whole genome gene expression arrays) in MPM subgroups with higher and lower sensitivity against trabectedin dichotomized by the median IC50 value (left panel) and linear regression analysis of the association between bcl-2 mRNA expression and trabectedin IC50 values (right panel). *, Student’s t test, p < 0.05. B, Ingenuity Pathway Analysis (IPA) indicated deregulation of a bcl-2-related module associated with intrinsic trabectedin responsiveness of MPM cell models. Expression differences (log fold change) are indicated below the gene symbols. Red and green indicate enhanced and reduced expression in the less responsive subgroup. Hoda et al

A P31 P31 res1.2 B Trabectedin (nM) Obatoclax (nM) - 1 2.5 5 - - 1 2.5 5 1 2.5 5 - - - - 250 500 250 250 250 500 500 500 - 1 2.5 5 - - 1 2.5 5 1 2.5 5 - - - - 250 500 250 250 250 500 500 500 PARP Cl. PARP Bim Bcl-2 Bcl-xL Bax β -actin P31 P31 res.1.2 Supplementary Figure S5. A, Activity of trabectedin combined with obatoclax against P31 cells and the cisplatin-resistant subline P31res1.2. After a 72 hours drug exposure, changes in viability were measured by MTT assay (left panels) and the respective combination indices (CI) were calculated (right panels). CI values <0.9, from 0.9 to 1.1, or >1.1 represent synergism, additive effects, and antagonism, respectively. B, P31 and P31res1.2 cells were exposed to increasing concentrations of trabectedin and obatoclax alone or in combination for 48 hours. Cleavage of PARP as well as expression of several key apoptosis-regulating proteins was analyzed by Western blot analysis. Hoda et al

SPC111 SPC212 P31 P31 res.1.2 Supplementary Figure S6. Activity of trabectedin combined with ABT-199 against MPM cell lines. Cells were continuously exposed to the indicated doses of trabectedin without or with ABT-199 for 72 hours. Changes in viability were measured by MTT assay and data are shown as dose response curves (left panels) and as combination indices (CI, right panels). CI values <0.9, from 0.9 to 1.1, or >1.1 represent synergism, additive effects, and antagonism, respectively. Hoda et al