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Journal: bioRxiv
Article Title: hENT Inhibition Prevents Pyrimidine-Driven Resistance to DHODH Inhibition in Malignant Rhabdoid Tumors
doi: 10.64898/2026.01.25.701565
Figure Lengend Snippet: A) Bar graph depicting the average viability (%) of ecMRT tumoroids cultured in KOM supplemented with 30 μM uridine following 120-hour treatment with various concentrations of dipyridamole. Data represent the mean ± SD of n = 3 independent experiments, each consisting of technical triplicates. Viability values were normalized to the DMSO vehicle control (100%). B) Dose-response curves for ecMRT tumoroid models 60T and 78T treated with GTX-196 for 120 hours in Plasmax™ medium, with or without the hENT1/2 inhibitor dipyridamole (500 nM). ecMRT tumoroids treated with GTX-196 in KOM medium served as a reference. Data represent mean ± SD from three independent experiments, each performed with technical duplicates. Data are normalized to DMSO vehicle in KOM (100%). The grey dotted horizontal line represents a viability of 50% (IC 50 ). C-D) Dose-response curves for ecMRT tumoroid models 60T treated with BAY-2402234 (panel C ) and 103T treated with GTX-196 (panel D) for 120 hours in KOM supplemented with 3 μM uridine, with or without different concentrations of hENT1/2 inhibitor dipyridamole. Data represent mean ± SD from three independent drug matrix experiments. Data are normalized to DMSO vehicle (100%). The grey dotted horizontal line represents a viability of 50% (IC 50 ). E-F) Two-dimensional (2D) synergy landscapes visualized as contour plots, displaying ZIP (Zero Interaction Potency) synergy scores across concentration matrices for BAY-2402234 combined with dipyridamole (panel E ) and GTX-196 combined with dipyridamole (panel F ) (corresponding to figure panels C and D ). ZIP scores >10 indicate synergistic interactions, scores between -10 and 10 represent additive effects, and scores <–10 indicate antagonism. Regions outlined in white represent the most synergistic concentration combinations. Synergy landscapes were generated using SynergyFinder software . G , I) Dose-response curves for ecMRT tumoroid model 103T treated with GTX-196 for 120 hours in KOM supplemented with 30 μM uridine, with or without different concentrations of hENT inhibitors draflazine (panel G ) or nitrobenzylthioinosine (NBMPR; panel I). Data represent mean ± SD from three independent drug matrix experiments. Data are normalized to DMSO vehicle (100%). The grey dotted horizontal line represents a viability of 50% (IC 50 ). H , J ) Two-dimensional (2D) synergy landscapes visualized as contour plots, displaying ZIP (Zero Interaction Potency) synergy scores across concentration matrices for GTX-196 combined with draflazine (panel H ) and GTX-196 combined with NBMPR (panel J ) (corresponding to figure panels G and I ). ZIP scores >10 indicate synergistic interactions, scores between -10 and 10 represent additive effects, and scores <–10 indicate antagonism. Regions outlined in white represent the most synergistic concentration combinations. Synergy landscapes were generated using SynergyFinder software . K) Isotopic labeling pattern of extracellular uridine levels measured in the medium of BME (no cells) and two different ecMRT tumoroids following 24-hour incubation with 3⍰μM or 30⍰μM [ 13 C 9 - 15 N 2 ]-uridine upon 48-hour treatment with DMSO vehicle, 5 nM GTX-196, 10 μM DP, or the combination thereof. All conditions were statistically compared to the BME (no cells) reference group. (*, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001).
Article Snippet: BAY-2402234 (MedChemExpress, HY-112645), GTX-196 (Genase Therapeutics), AG-636 (MedChemExpress, HY-137463), ASLAN003/Farudodstat (MedChemExpress, HY-129239), PTC299 (MedChemExpress, HY-124593), Dipyridamole (MedChemExpress, HY-B0312),
Techniques: Cell Culture, Control, Concentration Assay, Generated, Software, Isotopic Labeling, Incubation