Extended Data Fig. 3: Manipulation of Clu and Cd38 levels in HSCs affect differentiation of oHSCs in vivo.

(a) FACS profile and cell populations in the peripheral blood derived from HSCs infected with lentivirus after 4 months of transplantation in the recipient mice (top panel). FACS profile and cell populations of HSPCs derived from HSCs infected with lentivirus after 4 months of transplantation in the recipient mice (bottom panel). (b) Validation of Clu KO (3 sgRNA simultaneously targeting Exon3-5) in oHSCs by RNA-seq (left panel). Validation of Clu KO in oHSCs (middle panel) and Clu OE in yHSCs (right panel) by qPCR. (c) Leukocyte chimerism in the peripheral blood over time after competitive transplantation in the recipient mice with KO or OE of Clu in HSCs (n = 4 per group). (d) Frequency of lymphoid, myeloid, B, and T cells in mCherry⁺ peripheral blood cells at different time points post-transplantation in recipient mice with Cd38 KO oHSC (n = 5 per group). (e) Representative FACS plot (left) and quantification (right) of CD4⁺/CD8⁺ DP T cells in the thymus of recipient mice with control (sgCtrl) or Clu KO (sgClu) oHSCs (n = 4 per group). (f) Representative FACS plot (left) and quantification (right) of CD4⁺/CD8⁺ DP T cells in the thymus of recipient mice with control or Clu OE yHSCs (n = 4 per group). Unpaired two-tailed Student’s t-test (b–f). Error bars represent mean ± SEM (b, e, f). Error bars represent mean ± SD (c, d). Ns: no significant difference.

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