Ascorbate deficiency increases the reconstituting potential of bone marrow cells upon transplantation into irradiated mice. (A-B) Ascorbate levels in HSCs, MPPs, HPCs, and whole bone marrow cells (A) and blood plasma (B) measured by mass spectrometry (a total of 5-7 mice per genotype in 2 independent experiments). (C) Kaplan-Meier survival curve of Slc23a2-deficient and littermate control mice (a total of 8 mice per genotype in 3 independent experiments). (D-V) Bone marrow cellularity from 2 tibias and 2 femurs (D), as well as the frequencies of HSCs (E), MPPs (F), restricted hematopoietic progenitors (G-N), and differentiated B cells (O,S), T cells (P,T), neutrophils (Q,U), and monocytes (R,V) in the bone marrow (D-R) and blood (S-V) (a total of 6-12 mice per genotype from 6 independent experiments). (W-X) Donor cell reconstitution (W) in the blood and bone marrow (X) after transplantation of 500 000 donor bone marrow cells from Slc23a2-deficient or littermate control mice, along with 2 000 000 competing wild-type cells into irradiated recipients (a total of 13-14 recipient mice per genotype transplanted with cells from 3 donors per genotype in 3 independent experiments). All data are presented as the mean ± standard deviation. Each dot in panels A-V represents a different mouse. Statistical significance was assessed using Student t tests (D), Mann-Whitney tests (B and time points in W), log-rank tests (C), Mann-Whitney tests with Holm-Sidak’s correction for multiple comparisons (A,E-V,X), or nparLD nonparametric mixed models with Holm-Sidak’s multiple comparisons corrections to test differences in overall reconstitution (W). All tests were 2-sided (∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001).