Figure 6.

Decreased expression levels of ASXL1 in human leukemia cells are associated with AKT inhibitor sensitivity. (A and B) The inhibition ratios were measured by MTS assay after 48 h treatment of a concentration gradient of the AKT inhibitor MK2206 and the STAT3 inhibitor Stattics in Set2 cells (A) and HL60 cells (B). The IC₅₀ values were calculated accordingly. The error bars denote SD, n = 3. (C) Colony formation assay in methylcellulose was performed using primary bone marrow mononuclear cells isolated from MDS/CMML patients with (ASXL1-MT) or without (ASXL1-WT) ASXL1 mutation under treatment of the AKT inhibitor MK2206. CFU-GM was classified according to standard morphologic criteria. Data represent the counts of colonies from three independent experiments. (D) ASXL1-WT cells were transduced with two different shRNA constructs targeting ASXL1 (ASXL1 sh_C and ASXL1 sh_D) or scramble shRNA (Scr sh). CFU-GM assay was performed to assess the effect of the AKT inhibitor MK2206. Data represent the counts of colonies from three independent experiments. Statistical significance was determined by the two-tailed Student’s t-test. *P ≤ 0.05, **P ≤ 0.01, ****P ≤ 0.0001, NS indicates not significant.