Fig. 1

The collaboration of ASXL1 and RUNX1 mutants augmented proliferation and impair differentiation in vitro. a, b IL-3-dependent murine myeloid 32D cells stably transduced with ASXL1-R693X, RUNX1-WT/R135T, and combination of ASXL1-R693X and RUNX1-R135T were cultured in the absence (a) or in the presence (b) of IL-3 for 96 h. The number of viable cells is shown. c, d The stably transduced 32D cells were cultured with 50 ng/mL of G-CSF for 5 days; cell morphology with Wright–Giemsa-stained smear, original magnification × 400 (c), and percentage of CD11b-positive cells are shown (d). e, f The stably transduced U937 cells were cultured with 40 nM of PMA for 96 h; cell morphology with Wright–Giemsa-stained smears (original magnification × 400) (e, upper panel), representative flow cytometry analyses (e, lower panel), and percentage of CD11b-positive cells (f) are shown. All data error bars in a and b represented here are the mean ± SD of duplicate cultures and repeated three times; error bars in d and f represented the mean ± SD of four and three independent experiments respectively. *P < 0.05, **P < 0.03, ***P < 0.01; n.s., not significant