Figure 5.
Induction of cell death upon combined SUMO/proteasome inhibition is associated with PIN1. (A) OPM2 cells were treated for 4 hours with dimethyl sulfoxide (DMSO), 250 nM subasumstat, 5 nM CFZ, or the combination thereof and subsequently analyzed by RNA-sequencing. GSEA analysis by the fgsea package of the combination treatment (4 hours) vs DMSO control shows enriched p53 signatures of the Hallmark set from the molecular signature database. fgsea P values and adjusted P values (P-adj; false discovery rate) are indicated. (B) PIN1 SUMOylation upon 8 hours of 10 μM MG132 treatment or 1 hour of heat shock vs control with indicated P values. Data were retrieved from the qPTM database (http://qptm.omicsbio.info/) from the study. (C) Determination of viability of OPM2 cells, treated with 2.5 μM subasumstat or 1 μM sulfopin or combination of both for 72 hours. Cell viability was measured by CellTiterGlo. (D) Landscape plots depicting the antagonistic/additive or synergistic area of concentrations for subasumstat in combination with sulfopin treatment in OPM2 cells. Cells were treated for 72 hours with the indicated concentrations (4 × 6 matrix) of subasumstat and sulfopin and cell viability was measured by CellTiterGlo. Subsequently, cell viability data were used to generate landscape plots using SynergyFinder. (E) Immunoblots on OPM2 cell lysates treated for 4 hours with 250 nM subasumstat, 5 nM CFZ, 2 μM sulfopin, or the combination thereof. Protein expression of indicated proteins was detected using specific antibodies as indicated in the Material and methods section. Actin served as loading control. CL. Casp.3, cleaved caspase-3; Suba, subasumstat; Sulf, sulfopin; ZIP, Zero Interaction Potency method.

Induction of cell death upon combined SUMO/proteasome inhibition is associated with PIN1. (A) OPM2 cells were treated for 4 hours with dimethyl sulfoxide (DMSO), 250 nM subasumstat, 5 nM CFZ, or the combination thereof and subsequently analyzed by RNA-sequencing. GSEA analysis by the fgsea package of the combination treatment (4 hours) vs DMSO control shows enriched p53 signatures of the Hallmark set from the molecular signature database. fgsea P values and adjusted P values (P-adj; false discovery rate) are indicated. (B) PIN1 SUMOylation upon 8 hours of 10 μM MG132 treatment or 1 hour of heat shock vs control with indicated P values. Data were retrieved from the qPTM database (http://qptm.omicsbio.info/) from the study. (C) Determination of viability of OPM2 cells, treated with 2.5 μM subasumstat or 1 μM sulfopin or combination of both for 72 hours. Cell viability was measured by CellTiterGlo. (D) Landscape plots depicting the antagonistic/additive or synergistic area of concentrations for subasumstat in combination with sulfopin treatment in OPM2 cells. Cells were treated for 72 hours with the indicated concentrations (4 × 6 matrix) of subasumstat and sulfopin and cell viability was measured by CellTiterGlo. Subsequently, cell viability data were used to generate landscape plots using SynergyFinder. (E) Immunoblots on OPM2 cell lysates treated for 4 hours with 250 nM subasumstat, 5 nM CFZ, 2 μM sulfopin, or the combination thereof. Protein expression of indicated proteins was detected using specific antibodies as indicated in the Material and methods section. Actin served as loading control. CL. Casp.3, cleaved caspase-3; Suba, subasumstat; Sulf, sulfopin; ZIP, Zero Interaction Potency method.

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