Summary of competitive limiting dilution transplantation experiments
| Number of V617F;GFP+ or WT;GFP+ BM cells | Number of WT competitor BM cells | Ratio mutant/WT | WT;GFP+ | V617F;GFP+ | |
| Number and % of reconstituted mice | Number and % of reconstituted mice | Number and % of mice with MPN phenotype | |||
| 20,000 | 2,000,000 | 1:100 | 5/69 (7.2%)a | 15/40 (37.5%) | 4/15 (26.7%) |
| 8,000 | 2,000,000 | 1:250 | n.d. | 16/60 (26.7%) | 7/16 (43.8%) |
| 2,000 | 2,000,000 | 1:1000 | n.d. | 0/20 (0%) | 0/20 (0%) |
| Calculated frequency in total BMb | LT-HSC 1 in 265,000 | LT-HSC 1 in 35,000 | MPN initiating cell 1 in 113,000 | ||
| 20,000 | 1,000,000 | 1:50 | 5/30 (16.7%) | n.d. | n.d. |
| 8,000 | 1,000,000 | 1:125 | n.d. | 17/64 (26.6%) | 4/17 (23.5%) |
| 4,000 | 1,000,000 | 1:250 | n.d. | 3/20 (15%) | 1/3 (33.3%) |
| Calculated frequency in total BMb | LT-HSC 1 in 110,000 | LT-HSC 1 in 26,000 | MPN initiating cell 1 in 108,000 | ||
| Number of V617F;GFP+ or WT;GFP+ BM cells | Number of WT competitor BM cells | Ratio mutant/WT | WT;GFP+ | V617F;GFP+ | |
| Number and % of reconstituted mice | Number and % of reconstituted mice | Number and % of mice with MPN phenotype | |||
| 20,000 | 2,000,000 | 1:100 | 5/69 (7.2%) | 15/40 (37.5%) | 4/15 (26.7%) |
| 8,000 | 2,000,000 | 1:250 | n.d. | 16/60 (26.7%) | 7/16 (43.8%) |
| 2,000 | 2,000,000 | 1:1000 | n.d. | 0/20 (0%) | 0/20 (0%) |
| Calculated frequency in total BM | LT-HSC 1 in 265,000 | LT-HSC 1 in 35,000 | MPN initiating cell 1 in 113,000 | ||
| 20,000 | 1,000,000 | 1:50 | 5/30 (16.7%) | n.d. | n.d. |
| 8,000 | 1,000,000 | 1:125 | n.d. | 17/64 (26.6%) | 4/17 (23.5%) |
| 4,000 | 1,000,000 | 1:250 | n.d. | 3/20 (15%) | 1/3 (33.3%) |
| Calculated frequency in total BM | LT-HSC 1 in 110,000 | LT-HSC 1 in 26,000 | MPN initiating cell 1 in 108,000 | ||
The indicated numbers of BM cells were injected into lethally irradiated recipients. Reconstitution was scored based on the presence of >1% GFP-positive cells in the PB Gr1+ cells at ∼4 mo after transplantation. n.d., not done.
Numbers of mice from two separate experiments were pooled.
The frequencies of LT-HSC and MPN initiating cells were calculated by Poisson distribution based on the frequencies of reconstituted mice, and the frequencies of mice developing an MPN phenotype, respectively.
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