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S. Hultmark et al.
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Figure 2. H4 is a jatrophane with differentiation potential for primary FLT3 wild-type acute myeloid leukemia cells. (A) Molecular structure of the natural compound H4. (B) The optimal concentration of H4 was 10 mM for primary AML-3 cells. The data are shown as mean ± standard error of mean from independent experiments, DMSO (n=5), H4 5 mM (n=2), H4 10 mM (n=6), H4 20 mM (n=2), H4 40 mM (n=1) (**P<0.01, *P<0.05, ns not significant, using one-way analysis of variance). On the right, representative change in cell morphology and myeloid differentiation markers after treatment with H4 10 mM. Scale bars represent 10 μm. (C) No detected increase in the percentage of viable, apoptotic and necrotic AML-3 cells after treatment with H4 μM. The data are shown as mean ± standard deviation (SD) of tech- nical replicates. (D) Long-term culture of AML-3 cells on OP9M2 stroma cells. The data are shown as mean ± SD of technical replicates. (E) H4 induced differentiation of AML-3 cells that had been pretreated for 5 days with the CDK4/6 cell cycle inhibitor palbociclib, after 4-day combination treatment with the CDK4/6 cell cycle inhibitor palbociclib 5 mM. The data are shown as mean ± SD of technical replicates. (ns not significant, using an unpaired two-tailed t-test). (F) Engraftment levels of in vitro treated human AML3 cells transplanted into mice. (G) H4 triggered the strongest differentiation response in FLT3 wild-type acute myeloid leukemia sam- ples. The data are shown with the average response from each sample labeled with their respective sample number for easier identification. (significance test using an unpaired two-tailed t-test with the Welch correction). DMSO: dimethylsulfoxide; AML: acute myeloid leukemia; Ctrl: control; ITD: internal tandem duplication; mut: mutated.
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haematologica | 2021; 106(10)