Journal: Journal of Translational Medicine

Article Title: Targeting epigenetic features in clear cell sarcomas based on patient-derived cell lines

doi: 10.1186/s12967-022-03843-4

Figure Lengend Snippet: PRMT inhibitor treatment and ArgMet profiles: viability determination of MUG Lucifer prim and MUG Lucifer met cells treated with 0–10 µM GSK 3368715 for six days normalized to the DMSO control. IC 50 was calculated using GraphPad Prism 9. Error indicators are the positive and negative standard deviations based on biological replicates (n = 3) ( A ). Viability determination of MUG Lucifer prim and MUG Lucifer met cells treated with 0–10 µM GSK 591 for six days normalized to the DMSO control. Error indicators are the positive and negative standard deviations based on biological replicates (n = 3) ( B ). Viability determination of MUG Lucifer prim and MUG Lucifer met cells treated with 0–20 µM AdOx for 72 h normalized to the DMSO control. IC 50 was calculated using GraphPad Prism 9. Error indicators are the positive and negative standard deviations based on biological replicates (n = 3) ( C ). ADMA/ARG ratios of MUG Lucifer prim cells treated with 3.8 µM GSK 3368715 (IC 50 ) and 6.8 µM AdOx (IC 50 ) and the respective DMSO controls ( D ). ADMA/ARG ratios of MUG Lucifer met cells treated with 3.8 µM GSK 3368715 (IC 50 ) and 6.8 µM AdOx (IC 50 ) and the respective DMSO controls ( E ). (SDMA + MMA)/ARG ratios of MUG Lucifer prim cells treated with 3.8 µM GSK 3368715 (IC 50 ) and 6.8 µM AdOx, (IC 50 ) and the respective DMSO controls ( F ). (SDMA + MMA)ARG ratios of MUG Lucifer met cells treated with 3.8 µM GSK 3368715 (IC 50 ) and 6.8 µM AdOx (IC 50 ) and the respective DMSO controls ( G )

Article Snippet: The cells were treated with GSK 3368715 (MCE MedChemExpress, Monmouth Junction, NJ, USA), an uncompetitive type I PRMT inhibitor, in a concentration range of 0–10 μM for up to six days; with GSK 591 (MCE MedChemExpress), a potent and selective inhibitor of PRMT5, in a concentration range of 0–10 μM for up to six days; and with AdOx in a concentration range of 0–20 μM for up to three days in a humidified 5% CO 2 atmosphere at 37 °C; appropriate negative controls were used.

Techniques: Control

Journal: Journal of Translational Medicine

Article Title: Targeting epigenetic features in clear cell sarcomas based on patient-derived cell lines

doi: 10.1186/s12967-022-03843-4

Figure Lengend Snippet: Viability and apoptosis assays in 2D and 3D cell culture. 2D fluorescence/luminescence-based viability, cytotoxicity and apoptosis determination of MUG Lucifer prim cells treated with 3.8 µM GSK 3368715 (IC 50 ) for six days and MUG Lucifer met cells treated with 6.8 µM AdOx (IC 50 ) for 72 h including the respective DMSO and medium controls ( A ). Early apoptosis determination of MUG Lucifer prim cells treated with IC 50 + 20% GSK 3368715 (4.56 μM) for six days and MUG Lucifer met cells treated with IC 50 + 20% AdOx (8.12 μM) for 72 h in 2D cell culture using Annexin V/PI staining for flow cytometry. Annexin- PI + represent necrotic cells, Annexin + PI- cells represent early apoptotic cells, Annexin + PI + represent late apoptotic cells and Annexin- PI- cells represent viable cells ( B ). Viability determination of MUG Lucifer prim, MUG Lucifer met, MUG Lucifer hTERT fibroblasts and co-cultures of MUG Lucifer prim with MUG Lucifer hTERT fibroblasts (1 + 1 ratio) and MUG Lucifer met with MUG Lucifer hTERT fibroblasts (1 + 1 ratio) spheroids treated with IC 50 + 20% GSK 3368715 (4.56 μM) for six days normalized to the respective DMSO control (DMSO control = 100% viability) using CellTiter-Glo ® 3D Cell Viability Assay ( C ). Apoptosis determination of MUG Lucifer prim, MUG Lucifer met, MUG Lucifer hTERT fibroblasts and co-cultures of MUG Lucifer prim with MUG Lucifer hTERT fibroblasts (1 + 1 ratio) and MUG Lucifer met with MUG Lucifer hTERT fibroblasts (1 + 1 ratio) spheroids treated with IC 50 + 20% GSK 3368715 (4.56 μM) for six days normalized to the respective DMSO (DMSO control = 0% apoptosis) control using Caspase-Glo ® 3/7 Assay ( D ). Error indicators are the positive and negative standard deviations based on biological replicates (n = 3) ( A – D ). Spheroids of MUG Lucifer cell lines treated with IC 50 + 20% GSK 336815 (4.56 μM) for six days (right) and the respective DMSO controls (left). Scale bars represent 100 μm ( E )

Article Snippet: The cells were treated with GSK 3368715 (MCE MedChemExpress, Monmouth Junction, NJ, USA), an uncompetitive type I PRMT inhibitor, in a concentration range of 0–10 μM for up to six days; with GSK 591 (MCE MedChemExpress), a potent and selective inhibitor of PRMT5, in a concentration range of 0–10 μM for up to six days; and with AdOx in a concentration range of 0–20 μM for up to three days in a humidified 5% CO 2 atmosphere at 37 °C; appropriate negative controls were used.

Techniques: Cell Culture, Fluorescence, Staining, Flow Cytometry, Control, Viability Assay, Caspase-Glo Assay