Fig. 7.

Fig. 7.

HSPCs derived from alas2- and alad-deficient embryos show low erythroid lineage reconstitution efficiency. (A) Schematic workflow for the primary and secondary HSPC transplantation with sorted ubi:dsRed⁺/CD41:GFP^low HSPCs in alas2 and alad morphants at 5 dpf. (B) Flow cytometric analysis of engrafted ubi-dsRed⁺ cells in the recipient kidney marrow (KM) at 45 dpt. Number of recipients: n (control HSPCs>irradiated WT)=8, n (alas2-morphant HSPCs>irradiated WT)=6, n (alad-morphant HSPCs>irradiated WT)=7. (C) Statistical analysis of the engraftment efficiency in B; the total numbers of recipients are indicated. (D) Flow cytometric analysis of different blood cell types in recipient kidney marrows. Each circle and percentile represent different cell lineages (I, erythrocytes; II, myelocytes; III, lymphocytes; IV, progenitors). (E) Quantification of different blood lineages in the kidney marrows based on D. Total cell numbers in whole kidney marrows are indicated. (F) Statistical analysis of the number of RBCs in recipient peripheral blood. The total numbers of recipients are indicated. (G) Statistical analysis of the engraftment efficiency of control, alas2 and alad morphants after secondary transplantation. Number of recipients: n (control HSPCs>irradiated WT)=7, n (alas2-morphant HSPCs>irradiated WT)=7, n (alad-morphant HSPCs>irradiated WT)=5. Total numbers of recipients are indicated. (H) Quantification of different blood lineages in the kidney marrows based on G. Total numbers of recipients are indicated. Number of samples are indicated. Data are mean±s.d. **P<0.01, ***P<0.001 [one-way ANOVA, Tukey's multiple comparisons (C,F,G); two-way ANOVA, Sidak's multiple comparisons (E,H)]. n.s., not significant.