Extended Data Fig. 4 |. H3.3 maintains hematopoietic homeostasis and HSC repopulation.

a. Percentages of CD45.2⁺ cells at the week 0 and week 4 after tamoxifen injection; note the absence of DKO-CRA cells in the PB of recipients CD45.1⁺ recipients before tamoxifen induced deletion of H3.3A gene, suggesting a disadvantage of BKO HSPCs to repopulate the wild type BM in a competitive setting. Experiments were repeated twice; each with n=4 animals for each group. b. Survival probability of mice with different transplantation schemes. c. Representative flow cytometric plots for CD16/32⁺ LKS cells in the recipients transplanted with DKO or AKO BMMNCs. d. Percentages of B220⁺ B cells and CD11b⁺Gr1⁺ granulocytes in the spleen of recipients transplanted with DKO or AKO BMMNCs. e. Percentages of B220⁺ B cells, Gr1⁺ myeloid cells, and CD3⁺ T cells in the PB of recipients transplanted with DKO or AKO BMMNCs. f-g. Images showing the skin lesions of the DKO BMMNCs-transplanted recipient animals, at week 7 after H3.3A deletion. The frequency, onset, and treatment for such skin lesion is listed in g. h-k. Hematoxylin and eosin (H and E) staining of BM, spleen, liver, and lung of recipients transplanted with DKO or AKO BMMNCs (n=3 for DKO, n=4 for AKO). For the BM, note the hypocellularity and sclerosis at the epiphysis and metaphysis regions. For the spleen, note the purple pro-fibrotic fibrous structures and disappearances of white pups in the DKO mice. There was no apparent hematopoietic cell infiltration into the liver for DKO BMMNC cells transplanted mice. For a, p-value is calculated using two-way ANOVA. For panels d, e, the number of dots indicates the number of independent animals. Error bars indicate standard error of mean. For d, e, p-value is calculated using unpaired, two-tailed t-test; the number of dots equals to the number of biological replicates in d, e. Numerical source data are provided in Source Data.