Figure 4

Figure 4

Mice lacking Slc39a10 in hematopoietic cells have defective hematopoiesis and die within 1 day of birth. A) Alignment of the amino acid sequences of zebrafish (Danio rerio) slc39a10, mouse (Mus musculus) Slc39a10, and human (Homo sapiens) SLC39A10 proteins, as well as the scoring for protein homology obtained using ClustalW. B) Slc39a10 mRNA levels were measured in FL LT‐HSCs obtained from control and Slc39a10 cKO mice (n = 5 mice per group). C) Representative images of newborn (P1) control and cKO mouse pups. D) Kaplan–Meier survival curve of control and cKO mice (n = 8 and 10 for control and cKO, respectively). E) Example images of peripheral blood taken from control and cKO mice. F–H) Summary of red blood cell (RBC; F), white blood cell (WBC; G), and platelet (PLT; H) counts in the peripheral blood of control and cKO mice (n = 3 mice per group). I,J) Wright–Giemsa‐stained peripheral blood smears (I) and bone marrow smears (J) taken from control and cKO mice. Scale bars: 100 µm. K) Gross appearance of the whole body (top) and fetal liver (bottom) of control, heterozygous (het), and homozygous cKO embryos at E14.5. L) Absolute numbers of liver cells in control and cKO embryos measured at E14.5 (n = 10 and 7 for control and cKO, respectively). M) Fetal liver cells were obtained from E14.5 embryos, and long‐term repopulating HSCs (LT‐HSCs) were gated using flow cytometry. N,O) Summary of the percentage and absolute number of LT‐HSCs in the liver of control and cKO embryos (n = 10 and 7 for control and cKO, respectively). P) Schematic depiction of hematopoiesis showing the proliferation and differentiation of HSCs in the fetal mouse liver. Q,R) Gating strategy (Q) and absolute numbers (R) of the indicated erythrocyte developmental stages in the liver of control and cKO embryos were measured at E14.5 (n = 4 and 3 for control and cKO, respectively). S–U) Gating strategy (S), frequency (T), and absolute numbers (U) of myeloid cells (defined as Mac1⁺Gr1⁺), B cells (defined as CD19⁺), and T cells (defined as CD3⁺) in the liver of control and cKO embryos measured at E14.5 (n = 4 and 3 for control and cKO, respectively). Data in this figure are represented as mean ± SD. p values of survival in (D) were determined using the Log‐rank test, in (B), (F), (G), (H), (L), (N), (O), (R), (T), and (U) using 2‐tailed unpaired Student's t‐test. *p < 0.05, **p < 0.01, ***p < 0.001, and ns, not significant.