Fig. 6: Selinexor inhibits cytoplasmic localization and chromatin binding of NPM1-fusions.
From: NPM1-fusion proteins promote myeloid leukemogenesis through XPO1-dependent HOX activation
293 T cells were transfected with Flag-tagged NPM1::MLF1 (A) or Flag-tagged NPM1::CCDC28A (B) and were treated with vehicle (DMSO) or Selinexor (100 nM) for 24 h. Cells were stained with anti-Flag antibody. Cell nuclei were visualized with DAPI. Representative images (left) are shown. Scale bar, 10 μm. Subcellular localization of NPM1::MLF1 and NPM1::CCDC28A was evaluated in 100 cells (right). Mouse AML cells expressing NPM1::MLF1 (C) or NPM1::CCDC28A (D) were treated with 100 nM selinexor for 7 days. Binding of NPM1::MLF1 (C) and NPM1::CCDC28A (D) on the HOX gene cluster was assessed by ChIP-qPCR. Data are shown as mean ± SEM from triplicate wells. Two-tailed unpaired t test was used for pairwise comparisons. E 293 T cells were cotransfected with vector, MLL::ENL, NPM1::MLF1, NPM1::CCDC28A, wild-type NPM1 or NPM1c together with pGL4.74 vector and HoxA9 reporter. Selinexor was added 24 h after transfection (right), and the HOXA9 activation was measured 48 h after transfection using the Dual-Luciferase Reporter Assay System. Data are shown as means ± SEM from triplicate wells. One-way ANOVA was used for multiple comparisons.
