Figure 5. DEG-35 has preferential cytotoxic activity in AML cells compared to normal cells.

(A) Normalized colony assay of wt LSK cells, Crbn^I391V LSK cells, MLL-AF9 wt cells and MLL-AF9 Crbn^I391V cells treated with different concentrations of DEG-35 plated in M3434 methylcellulose for 5 days. Cells from n=3 mice for wt LSK cells, Crbn^I391V LSK cells, MLL-AF9 Crbn^I391V and n=2 mice for MLL-AF9 wt cells. Data shown is the mean ± SEM of the different biological replicates tested from two different experiments. (B) Viability assay of wt LSK, Crbn^I391V LSK, MLL-AF9 wt and MLL-AF9 Crbn^I391V cells treated with DEG-35 for 3 days. Data shown is the mean ± SEM of three different biological replicates tested. (C) Normalized colony assay of eight primary AML patient cells and four normal bone marrow or CD34+ HSPC cord blood cells treated with DEG-35 plated in H443 methylcellulose for 7 days. The mean ± SEM of indicated samples tested from more than three different experiments is shown. (D) Viability assay of AML PDX cells and normal BM cells treated with DEG-35 for 2 days. Data shown is the combined result of two experiments using normal BM (n=2) and PDX cells (n=2). Mean ± SEM of triplicates. (E) Western blot analysis of AML PDX sample 1 treated with different concentration of DEG-35 for 24h. For results in (A, C), Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001. See also Figure S5.