Fig. 5

WNT3 silencing modulates the growth and imatinib mesylate response of CML cells. A Two independent shRNA sequences against WNT3 were delivered into K562 cells with lentiviral vectors. The transcript expression of WNT3 in shRNA-transduced and control cells were analyzed by RT–qPCR. The total lysate and secretion fraction of shRNA-transduced and control cells was analyzed by western blotting. Ponceau S staining was used to monitor the sample loading of the secreted proteins. B, C The growth and colony-forming cell (CFC) production of WNT3-silenced and control cells were analyzed (n = 3). D WNT3-silenced and control cells were plated for the CFC assay with or without imatinib mesylate (IM) (n = 3), and the percentage of surviving CFCs was calculated and compared (n = 3). E The expression of WNT3 in shWNT3-transduced and control CML CD34⁺ cells was analyzed by RT–qPCR (n = 3). F CFC productions of WNT3-silenced and control CML CD34⁺ cells (n = 4) were compared. BFU-E, burst-forming unit-erythroid; CFU-GM, colony-forming unit-granulocyte/macrophage; Mix, colony-forming unit-granulocyte, erythroid, macrophage, megakaryocyte. G WNT3-silenced and control CML CD34⁺ cells (n = 5) were plated for CFC assays with or without IM. 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, ***P < 0.001, and ****P < 0.0001)