A. RPMI8226, MM.1S and LP1 cells were treated with or without GSK126 (15 μM) for 24 h, and then ALDH+ cells were examined using ALDEFLUOR™ Kit (STEMCELL Technologies). DEAB was used as negative control. Representative dot plots (top) and statistical analysis (bottom) of 3 independent experiments are shown. B. LP1 cells were treated with or without GSK126 (15 μM) for 24 h, and the side population (SP) cells were detected by staining with Heochst33342 (5 μg/ml) for 90 min. Veraparmil (50 μM) was used as negative control. C-D. After transfected with vector, EZH2-WT and EZH2-H694A plasmids, respectively, LP1 cells were subjected to ALDH (C) and SP (D) assay. E. RPMI8226, MM.1S and LP1 cells exposed with indicated concentrations of GSK126. Regulatory proteins in Wnt/β-catenin pathway were analyzed by immunoblotting. F. After treatment with different concentrations GSK126 for 24 h, the mRNA expression levels of LEF1, c-Myc in LP1 cells were detected by qRT-PCR. GAPDH was used as a reference gene. G-H. After transfected with vector, EZH2-WT and EZH2-H694A plasmids, respectively, LP1 cells were subjected to immunoblotting (G) and qRT-PCR (H) analysis. I-J. After LP1 cells were treated with different concentrations GSK126 for 24 h (I) or transfected with vector, EZH2-WT and EZH2-H694A plasmids (J), respectively the mRNA expression levels of CXXC4, NKD1 and PRICKLE1 were detected by qPCR *, P < 0.05; **, P < 0.01, Student's t test. Arrows indicates the specific bands of corresponding proteins.