Enhanced hematopoietic reconstitution caused by loss of PHF6 is due to increased production of differentiated progeny. (A) Experimental design for transplants of each indicated population. (B) Contribution of CD45.2⁺ HSCs to peripheral blood. (C) Contribution of CD45.2⁺ MPP cells to peripheral blood. (D) Self-renewal of transplanted CD45.2⁺ HSCs, as calculated by dividing the final number of CD45.2⁺ HSCs per femur primary transplants by the input (100). (E) Differentiation of transplanted CD45.2⁺ HSCs to WBCs, calculated by dividing the final number of CD45.2⁺ WBCs per microliter of blood by the final number of CD45.2⁺ HSCs per femur. (F) Differentiation of transplanted CD45.2⁺ HSCs to progenitor populations, calculated by dividing the final number of each CD45.2⁺ progenitor population per femur by the final number of CD45.2⁺ HSCs per femur. (G) Representative images of spleens 12 days posttransplantation of 100 control or Phf6^lox/Y;Tie2-cre^Tg/+ HSCs. Scale bars, 1 cm. (H) Number of colonies counted on spleens and spleen weight (divided by host mouse weight) 12 days posttransplantation of control or Phf6^lox/Y;Tie2-cre^Tg/+ HSCs. (I) Images showing example of symmetric self-renewal (low Numb expression), symmetric differentiation (high Numb expression), and asymmetric differentiation (asymmetric Numb distribution). Scale bars, 2 µm. (J) Frequency of symmetric self-renewing, symmetric differentiating, and asymmetric divisions, as determined by Numb staining in control vs Phf6^lox/Y;Tie2-cre^Tg/+ HSCs, cultured for 48 hours, with addition of the mitosis inhibitor nocodazole after 24 hours. n = 4 control (3 Phf6^+/Y;Tie2-cre^Tg/+, 1 Phf6^+/Y), n = 4 Phf6^lox/Y;Tie2-cre^Tg/+ donors in panels A-F. n = 3 control (Phf6^+/Y;Tie2-cre^Tg/+), n = 3 Phf6^lox/Y;Tie2-cre^Tg/+ donors in panels G-H. n = 4 control (2 Phf6^+/Y;Tie2-cre^Tg/+, 1 Phf6^+/Y, 1 Phf6^lox/Y), n = 4 Phf6^lox/Y;Tie2-cre^Tg/+ mice in panels I-J. All donors were 12 weeks old. Data in panels B-C were arcsine transformed prior to being analyzed by the 2-tailed Student t test and are displayed as mean ± SEM. Data in panels D-F,H were analyzed by the 2-tailed Student t test and are mean ± SEM, with individual data points shown for each transplant recipient. Data in panel J were analyzed by the 2-tailed Student t test and are mean ± SEM.