Fig. 2

Functional characterization of cell cycle-restricted pre-leukemic stem cells (pre-LSCs). a Scheme for serial transplantation into primary (I), secondary (II), tertiary (III) and quaternary (IV) recipients, using purified GFP^lo and GFP^hi DN3 thymocytes. b Fold expansion of donor-derived DN3 thymocytes enumerated in the thymus of primary recipient mice. Values are median ± 95% confidence interval (CI), Student’s t-test, n = 15. c Immunophenotype of donor-derived cells in the thymus of primary recipients. DN, DP and SP represent the CD4⁻CD8⁻ double-negative, CD4⁺CD8⁺ double-positive and CD4⁺CD8⁻ single-positive with CD4⁻CD8⁺ single-positive populations, respectively. Values are mean ± s.d., Student’s t-test, n = 5. d Fold expansion of donor-derived DN3 thymocytes in the thymus of recipients serially injected with either GFP^lo or GFP^hi DN3 Lmo2^Tg thymocytes. The frequency of recipients ultimately developing donor-derived leukemia is indicated in the top gray shade diagram. Number of leukemic recipients over the total number of recipients injected per cohort is indicated. Values are mean ± s.e.m., Student’s t-test. e Scheme for green fluorescent protein (GFP) labeling of donor-derived DN3 thymocytes in primary transplants (top) and proportion of donor-derived GFP^hi DN3 cells (bottom, framed) in the thymus of primary recipients, 2 weeks post pulse. Values are mean ± s.d., Student’s t-test. f Proportion of chemoresistant DN3 thymocytes from each GFP-labeled subset found in H2B-GFP;Lmo2^Tg mice 2 weeks after doxycycline pulse. VXL induction-like therapy for T-ALL, including vincristine, dexamethasone and l-asparaginase²⁷. Values are mean ± s.d., Student’s t-test *p < 0.05, ***p < 0.001