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. 2022 Feb 26;8:85. doi: 10.1038/s41420-022-00877-x

Fig. 1. Ewing Sarcoma cells are sensitive to DDK inhibition.

Fig. 1

A Ewing Sarcoma cells (A673, RD-ES and SK-ES-1) and the non-Ewing, osteosarcoma cell line U2OS were treated with increasing concentrations of the DDK inhibitors XL413 and TAK-931 for 72 h and cell viability was measured using CCK-8 cell viability reagent. Relative viability was calculated based on DMSO treated cells (n = 3 biological replicates). 2-way ANOVA multiple comparisons, ***p < 0.001, ****p < 0.0001. B Cells were treated with either 0.1% DMSO, 5 µM XL413 (left panel) or 1 µM TAK-931 (right panel) for 48 h. Whole-cell lysates were collected, and western blot was performed for the specified proteins. All original western blot images are included in Supplementary Data. GAPDH was used as a loading control. C Ewing Sarcoma cells were treated with 0.1% DMSO, 5 µM XL413 or 1 µM TAK-931 for 48 h. Cells were stained with anti-Annexin-V (AlexaFluor-488 conjugated) and propidium iodide and analyzed using flow cytometry. Apoptotic cells were designated as Annexin-V (upper right and lower right quadrants) positive cells and were measured using FCS express v7 (n = 3 biological replicates). Bars represent the mean of the data. Unpaired t test, ***p < 0.001, **p < 0.01.