Fig. 2
Monitoring of cellular fitness and proliferation of MOLM-13 after treatment with AzaC or AzadC. a Brightfield microscopy images of MOLM-13 treated for 72 h with different concentrations of either AzaC or AzadC. Untreated cells served as a control. b MTT assay results to measure the metabolic activity after 48 h or 72 h exposure to 0.5 µM, 1.0 µM and 2.5 µM (increasing concentrations are indicated by the triangle) AzaC or AzadC. The metabolic activity of treated cells was individually normalized to the metabolic activity of the untreated control for every independent experiment. Each dot represents one independent experiment. Ordinary two-way ANOVA combined with Tukey’s multiple comparisons test was performed. All p values were adjusted for multiple comparisons testing. ns padj ≥ 0.05, *padj < 0.05, **padj < 0.01, ***padj < 0.001, ****padj < 0.0001. c Flow cytometric analysis of proliferation using EdU after 48 h of treatment with 0.5 µM, 1.0 µM and 2.5 µM of AzaC or AzadC. On the x-axis the signal intensity of the EdU, which has been conjugated to Alexa488 by click chemistry, is displayed for each detected cell. The count (y-axis) displays how many cells with a certain EdU signal intensity have been detected. Cells that had an EdU signal intensity > 1.5 × 104 were considered to have completed S-phase and reached G2 phase (orange area), EdU intensity > 2 × 103, but < 1.5 × 104 was considered as S-phase (green) and EdU signal < 2 × 103 represents cells that had not entered S-phase within the two hours of EdU exposure (violet area). b Details about the analysis, including exact p values, are given in Additional file 12: Table S11