Fig. 4: Experimental validation of inferred pro-differentiation and randomly selected drugs in HL-60.

A Time line of the experiment to test the pro-differentiation activity of drugs on HL-60 cells. B Top 30 ranking compounds with corresponding experimental outcomes. Compounds are ranked according to their “pro-differentiation score”. Compounds that have shown or not a significant activity on HL-60 differentiation are labelled with orange or yellow background respectively. A compound that was toxic at the concentrations used in the assay is labelled with a grey background. C Quantitation of NBT assay of HL-60 treated with the drugs listed in Supplementary Figure 2A. All drug treatments were carried out for four days. The concentration for each drug is reported in Table 1 in Materials and Methods section. The NBT assay results were plotted as percentage of positive cells over the total cells. D Cell viability after four days of selected drug treatment. Cell viability was assessed by the Trypan blue exclusion test. E Representative light microscope images of NBT staining in HL-60 after four days of drug treatment. Scale bar is 50 μm. F Quantitation of the NBT assay of HL-60 treated with the randomly selected drugs listed in Supplementary Figure 2B. The concentration for each drug is reported in Table 1 in the Materials and Methods section. The NBT assay results were plotted as percentage of positive cells on the total cell number. G Dot plots of the results of NBT assays on HL-60 treated with randomly selected compounds (blue circles) or predicted compounds (red squares). Each circle or square represents the mean of the percentage of NBT positive cells over total cells of three biological replicates. Statistical significance was evaluated using a Student t-test (n = 3). Data are presented as mean ± SEM. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. Data in (C), (D) and (F) are represented as means of three biological replicates (n = 3) ± SEM. Statistical analysis was performed using One-way ANOVA. Significance *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 and are related.