Fig. 1

WT1 is required for the maintenance of MLL-AF9-induced murine leukemia. a The transcript expression of wt1 is measured in LICs (GFP⁺c-Kit⁺Mac-1⁺) and LSCs (GFP⁺Lin⁻c-Kit⁺Sca-1⁻CD34⁺CD16/32⁺) from MLL-AF9-induced murine leukemia (n = 4) and BM Lin⁻ cells from normal C57/B6 mice (n = 4). b A schematic outline of the in vivo experiment using MLL-AF9-induced murine AML model. c The frequency of GFP⁺ cells was measured in BM cells from MLL-AF9-induced leukemia with knockdown of wt1 (n = 4) or control nc (n = 4). Shown are the representative plots (left) and statistical analysis of the frequency of GFP⁺ cells (right). d A representative image of the spleen (left) and statistical analysis of spleen weight (right) in the MLL-AF9-induced murine leukemia with knockdown of wt1 (n = 4) or control nc (n = 4). e Frequency of LICs was measured in GFP⁺ BM blasts from MLL-AF9-induced leukemia with knockdown of wt1 (n = 4) or control nc (n = 4). Shown are the representative plots (left) and statistical analysis of the frequencies of c-Kit⁺Mac-1⁺ cells (right). f Percentage of L-GMP was assessed in BM GFP⁺ blasts from MLL-AF9-induced leukemia with knockdown of wt1 (n = 4) or control nc (n = 4). Shown are the representative plots (left) and statistical analysis of the frequencies of L-GMP (right). g BM GFP⁺ cells were isolated from MLL-AF9-induced leukemia with knockdown of wt1 (n = 4) or control nc (n = 4) and then plated on methylcellulose medium. Colonies were counted after 2 weeks (n = 4). Shown are the representative colony pictures (left) and statistical analysis of the number of colonies (right). Bar represents 50 µm. h–j Overall survival was determined in the primary BMT (h, n = 8), secondary BMT (i, n = 8), and tertiary BMT (j, n = 8) of MLL-AF9-induced murine leukemia with knockdown of wt1 or control nc. k Limiting dilution assay of BM GFP⁺ cells from MLL-AF9-induced murine leukemia with knockdown of wt1 (n = 6) or control nc (n = 6). The frequency of LSC and P-value were calculated by L-calc software