Fig. 3: IPO11 is necessary for primary AML stem/progenitor cells.

a Clonogenic growth of OCI-AML2, NB4 and TEX cells transduced with shRNA targeting IPO11 or control sequence. Mean ± SD colony counts are shown per ml (750 cells). *p ≤ 0.05, **p ≤ 0.01, by t-test. b TEX AML cells were transduced with shRNA targeting IPO11 or control sequences. 6 days after transduction, equal numbers of viable cells were injected into sub-lethally irradiated NOD/SCID-GF mice. The percentage of human CD45+cells in the non-injected femur was determined by flow cytometry (n = 8–9/group) after five weeks. The bar represents mean engraftment. p < 0.0001 by t-test. c TEX cells transduced with either non-target control or shRNA targeting IPO11 were injected into NOD.SCID-GF mice. Survival was measured over time. d 8227 cells were transduced with shRNA targeting IPO11 or control sequences in lentiviral vectors containing a GFP marker. Two days after transduction, equal numbers of viable cells were injected into the sub-lethally irradiated NOD/SCID-GF mice (n = 6 mice/group). Eight weeks later, the percentage of human GFP+, CD45+ cells in the non-injected femur was determined by flow cytometry. The transduction efficiency of the control and IPO11 shRNA lentiviral vectors were 30% and 32%, respectively. The bar represents mean engraftment, p < 0.0001 by t-test. e IPO11 expression in 8227 leukemia cells FACS sorted into bulk (CD34−) and stem cell (CD34+) fractions. f Primary AML cells were transduced with shRNA targeting IPO11 or control sequences in lentiviral vectors containing a GFP marker. Two days after transduction, equal numbers of viable cells were injected into sub-lethally irradiated NOD-SCID mice preconditioned with anti-CD122. Eight weeks after injection, the percentage of human GFP+, CD33+ and CD45+ cells in the non-injected left femur was determined by flow. g Engraftment of primary AML cells from relapse following IPO11 KD, same as described above. Transduction efficiency of NT control and IPO11 shRNA were 36% and 38% for patient 120878, 24% and 30% for patient 120858, 20% and 25% for patient 130414, respectively. IPO11 KD was confirmed by qPCR in GFP+ sorted cells. The bar represents mean engraftment. h Secondary engraftment of primary AML (130414) cells. Equal number of cells from mice in (g) were injected into the right femur of secondary untreated mice. Eight weeks after injection, the percentage of human GFP+, CD33+ and CD45+ cells in the non-injected left femur were determined by flow.