Fig. 5

Ouabain exhibits greater cytotoxicity in LSCs than normal HSPCs. A Primary AML cells from relapsed patients, including Pr3R (left) and Pr2R (right) cells, were similarly treated with ouabain (0 − 100 nM) for 48 h. (upper) Profiles of different AML subpopulations, including CD34⁺CD38⁻ LSCs, CD34⁺CD38⁺ LPCs, CD34⁻CD38⁺ pre-LBs, and CD34⁻CD38⁻ LBs, in primary AML cells in StemSpan culture by flow cytometry. (lower) Percentage of cell death (Annexin V⁺) in pooled Pr3R AML cells or enriched Pr2R LSCs and LBs. Data are mean ± SD (n = 3 or 5). ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 versus nontreated control; ^##P < 0.01, ^###P < 0.001 versus ouabain-treated LSCs at the same concentration; two-sided Student’s t test. B Enriched LSCs from AML KG-1 cells and enriched normal HSPCs from cord blood (CB) and G-CSF-mobilized peripheral blood were similarly treated with ouabain (0 − 100 nM) for 24 − 48 h. Cell death was evaluated by Annexin V/7-AAD assay and percentage of cell death was plotted. Data are mean ± SD (n = 3). ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 versus nontreated control; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 versus ouabain-treated LSCs at the same concentration; two-sided Student’s t test