Fig. 6

Anti-self-renewal effect of WP1130 on LSCs in MLL-AF9-induced murine leukemia. a A schematic outline of the in vivo experiment using MLL-AF9-induced murine leukemia model untreated or treated with WP1130. b GFP⁺ cells were measured in BM cells isolated from MLL-AF9-induced leukemic mice untreated (n = 4) or treated with WP1130 (n = 4) upon the development of full-onset leukemia by vehicle-treated mice. Shown are the representative plots (left) and statistical analysis of the frequency of GFP⁺ cells (right). c A representative image of blood and BM smear of vehicle- or WP1130-treated MLL-AF9 mice (left) and statistical analysis of the average percentage of leukemic blasts in blood and BM (right). Bar represents 10 µm, and these images were amplified 200 folds. ^** and ^##P < 0.01. d A representative image of the spleen (left) and statistical analysis of spleen weight (right) from the vehicle-(n = 4) or WP1130-treated mice (n = 4). e The frequencies of LICs in GFP⁺ cells were assessed in BM cells from vehicle- (n = 4) or WP1130-treated mice (n = 4). Shown are the representative plots (left) and statistical analysis of LICs (right). f Limiting dilution assay of BM GFP⁺ cells from vehicle- (n = 6) or WP130-treated mice (n = 6). The frequency of LSCs and the P-value were calculated by L-calc software. g BM GFP⁺ cells were isolated from vehicle- (n = 4) or WP1130-treated mice (n = 4) and then plated on methylcellulose medium. Colonies were scored after 2 weeks. Shown are the representative pictures showing colonies (left) and statistical analysis of the number of colonies (right). Bar represents 50 µm. (h–j) Overall survival was analyzed in the primary BMT (h, n = 8), secondary BMT (i, n = 8), and tertiary BMT (j, n = 8) of MLL-AF9-induced leukemia mice treated with vehicle or WP1130. (K) Wt1 and bcl2l2 protein expressions were measured in BM cells from vehicle- (n = 2) or WP1130-treated mice (n = 2)