Extended Data Fig. 4: ThPOK loss does not affect MDP development.

a) Bar graphs showing absolute cell numbers for CMP, GMP, MEP and CD34^lo subsets among gated cKit⁺ Sca1^- Lin^- BM cells from ThPOK-deficient or WT mice (same mice as Fig. 2a, b) (n = 5 independent animals per genotype). Error bars represent SEM. Significant differences between ThPOK^-/- and WT mice were determined by two-sided unpaired T test with Welch’s correction, and indicated by asterisks (* p < 0.05; ** p < 0.01; *** p < 0.001). b) FACS analysis of CD135 and CD115 expression by gated CMPs from WT and ThPOK^-/- mice (left panels). Bar graph at right shows % of indicated gated subsets (n = 5 independent animals per genotype). c) FACS analysis of CD11b, CD115, Ly6c, and Ly6g expression by gated cGMP (conventional GMP) (grey histograms) and c-Kit⁺ CD34^lo CD16^hi Lin^- (green histograms) BM subsets from WT mice. d) FACS analysis of CD16/32 and CD34 expression by gated Lin^- c-Kit⁺ Sca1^- BM cells from ThPOPK^+/- mice. CMP, GMP, MEP and CD34^lo subsets are marked. e) CFU assay of FACS-sorted Lin^- BM cells, from indicated WT or ThPOK-deficient mice. Error bars represent SEM. Significant differences between ThPOK^-/- and WT mice were determined by two-sided multiple multiple unpaired T with Welch’s correction, and indicated by asterisks (* p < 0.05; ** p < 0.01; *** p < 0.001). Note that ThPOK-/- Lin^- progenitors exhibited a substantial (** p < 0.01) increase in CFU-GM myeloid colony production even after secondary plating. f) FACS analysis of CD11b, Thy1, CD41 and Ter119 lineage marker expression by WT and ThPOK-deficient cells after secondary CFU assay (same experiment as in panel d).

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