Supplementary Figure 8: LAIR1 enhances leukemia development in several mouse leukemia models during serial transplantation.

(a) No significant difference in AML development were observed on primary transplantation with MLL-AF9-infected WT or lair1-null hematopoietic progenitors in primary transplantation; survival curves are shown (n = 10 mice; p = 0.1121, log-rank test). The experiment was repeated three times with similar results. (b) Summary of percentages of YFP⁺ AML cells, YFP⁺Mac-1⁺Gr-1⁺, and YFP⁺Mac-1⁺Kit⁺ cells in BM of primary recipient mice transplanted with the WT or lair1-null MLL-AF9 AML cells (n = 10 mice), the experiment was repeated three times with similar results. (c) The clonal relationship between the transplanted cells was studied by southern blotting on genomic DNA isolated from 4 pairs of WT and lair1 null MLL-AF9 induced leukemia samples. The result demonstrates that the leukemias were oligoclonal. The southern blotting was performed using a probe for YFP. (d) Southern blotting analysis of the oliogocolnal nature of MLL-AF9 integration in genomes of 3 primarily and 2 secondarily transplanted mice. The secondarily transplanted AML mice contained multiple MLL-AF9 clones from different primary samples. The bands indicated by the red number coordinates between primary and secondary samples. (e) Survival curves of mice receiving N-Myc infected WT or lair1-null hematopoietic progenitors in primary transplantation (n = 10 mice,p = 0.4204, log-rank test). (f) Survival curves of mice receiving 3,000 pooled GFP⁺ BM cells that were collected from primary recipients transplanted with WT or lair1-null N-Myc B-ALL cells (n = 5 mice, p = 0.0031, log-rank test). (g–i) Leukemia development vanishes on secondary transplantation of lair1-null MLL-AF9 cells. Summary of percentages of YFP⁺ AML cells and YFP⁺Mac-1⁺Kit⁺, YFP⁺Mac-1⁺Gr-1⁺, YFP⁺B220⁺, and YFP⁺CD3⁺ cells in (f) BM, (g) PB, and (h) spleen of secondary recipient mice transplanted with the WT or lair1-null MLL-AF9 AML cells at day 28 post-transplant (mean ± s.e.m., Student’s t-test; n = 5 mice; (g) Mac1⁺/Gr1⁺ **p = 0.0015; B220⁺ **p = 0.0019; CD3⁺ ***p < 0.0001; (h) Mac1⁺/Gr1⁺ **p = 0.0031; B220⁺ **p = 0.0022; CD3⁺ **p = 0.0017; (i) Mac1⁺/Gr1⁺ **p = 0.0002; B220⁺**p = 0.0035; CD3⁺ ***p < 0.0001).