Fig. 2

ZFX silencing inhibits CML CD34⁺ cells and sensitizes them to imatinib mesylate treatment. A Two independent shRNA sequences against ZFX were delivered into CML CD34⁺ cells (n = 3) or normal bone marrow (NBM) CD34⁺ cells (n = 3) with lentiviral vectors, and the relative expression of ZFX in shRNA-transduced cells compared with control (scramble) cells was measured by RT–qPCR. B The colony-forming cell (CFC) productions of ZFX-silenced and control cells were compared. BFU-E, burst-forming unit-erythroid; CFU-GM, colony-forming unit-granulocyte/macrophage; Mix, colony-forming unit-granulocyte, erythroid, macrophage, megakaryocyte. C ZFX-silenced and control CML CD34⁺ cells (n = 6) were plated for CFC assays with or without imatinib mesylate (IM). D Two independent shRNA sequences against Zfx were delivered into BaF3–BCR/ABL cells, and the relative expression of Zfx in shRNA-transduced cells compared with control cells was assessed by RT–qPCR. E, F The growth and CFC production of Zfx-silenced (shZfx) and control BaF–BCR/ABL cells were analyzed. G The CFC production of Zfx-silenced and control cells with or without IM was analyzed, and the percentage of CFC production relative to untreated cells was compared. H Zfx-silenced and control BaF–BCR/ABL cells were injected into lethally irradiated mice through the tail vein (seven mice in each group), and the survival of these mice was analyzed with the Kaplan–Meier method (log-rank test, **P < 0.01). I Leukemic cells were collected from the diseased mice of each group, and the expression of Zfx was analyzed by RT–qPCR and western blotting. J The weights of the liver and spleen from the diseased mice were measured, and the organ coefficient (organ weight/body weight, g/g) was calculated to evaluate the severity of leukemia symptom. Data are presented as the mean ± SEM, and Student’s t-test was used to estimate the P-values (*P < 0.05, **P < 0.01, and ***P < 0.001)