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. 2016 May 9;7(23):33512–33528. doi: 10.18632/oncotarget.9257

Figure 4. Combined treatment with L-asparaginase (Asp) and TRAIL results in anti-proliferative and pro-apoptotic synergism.

Figure 4

A., SF188 and LN299 glioblastoma cells were treated for 72h with L-asparaginase and TRAIL as indicated prior to performing MTT-assays. Normalized isobolograms were calculated using the CompuSyn software. The connecting line represents additivity. Data points located below the line indicate a synergistic drug-drug interaction and data points above the line indicate an antagonistic drug-drug interaction. B., Representative flow plots of SF188 (Asp 0.5IU/ml, TRAIL 25 ng/ml), LN229 (Asp IU/ml, TRAIL 100 ng/ml) and T98G (Asp 0.75IU/ml, TRAIL 4ng/ml) glioblastoma cells subjected to treatment with L-asparaginase, TRAIL or the combination for 24h prior to staining for annexin V/propidium iodide and flow cytometric analysis. C., SF188 and LN229 glioblastoma cells were treated with L-asparaginase and/or TRAIL as indicated. Western blot analysis was performed for caspase8 (CP8), cleaved caspase 3 (cCP3), cleaved PARP (cPARP), Mcl-1, Usp9X and Noxa. Actin expression was determined to confirm equal protein loading. D., Representative flow plots of T98G glioblastoma cells treated with increasing concentrations of L-asparaginase prior to staining for JC-1 and flow cytometric analysis. E, Representative microphotographs of LN229 glioblastoma cells treated with 2 IU/ml L-asparaginase, 250 ng/ml TRAIL or the combination for 48h. Magnification, x40; scale bar, 100 μm.