ADAMTS13 is required for proper hematopoietic lineage maturation in the adult zebrafish. (A-D) Flow cytometry analysis of kidney marrow-derived cells from WT (grey) and a13i5 (blue) adult zebrafish. (A) Gating strategy highlighting the main KM cell populations: cell debris (1), lympho-erythroid (2), myeloid (3) and early progenitors (4). Bar graphs represent the relative proportions of myeloid (B), lympho-erythroid (C), and progenitor (D) cell populations. (E-H) RT-qPCR analysis of lineage-specific gene expression in WT and a13i5 mutants. Expression of the macrophage markers mpeg1 (E) and marco (F), the neutrophil marker mpx (G) and the erythroid marker gata1 (H) normalized to the housekeeping gene ube2a and shown relative to the WT. Data are represented as mean±s.d. Each dot represents a biological replicate. Statistical analysis was performed using a two-tailed Student's t-test: *P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001. (I) Summary scheme of the effect of the lack of adamts13 on hematopoietic lineage specification. HSC, hematopoietic stem cell; CLP, common lymphoid progenitor; CMP, common myeloid progenitor; MP, macrophage; RBC, red blood cell; MK, megakaryocyte.
|
