Figure 1.
DPY30 Deficiency in the Fetal Hematopoietic System Results in Anemia and Defective HSC Function
Vav-Cre; Dpy30F/+ (F/+) and Vav-Cre; Dpy30F/− (F/−) littermate fetuses were used in (A–E and I).
(A) Image of littermate fetuses at E14.5 and E15.5.
(B) Relative Dpy30 mRNA levels in whole FL and sorted hematopoietic cell populations determined by qRT-PCR and normalized to Actb. n = 3–6 each for FL, n = 4 each for sorted cells.
(C) Immunoblotting for different levels of H3K4 methylation and other proteins in FL. Right, quantification from five embryos each.
(D) Absolute numbers of FL cell populations. n = 5 each.
(E) Colony formation assay using FL cells, showing representative images of colonies and quantification. n = 3 each.
(F) Scheme for the mixed chimera transplantation system using whole FL cells from Mx1-Cre; Dpy30F/+ or Mx1-Cre; Dpy30F/− fetuses as donors and whole BM cells from wild-type mice as competitors.
(G) Donor contribution to different cell populations in chimeras at indicated times after pIpC injections following scheme in (G). n = 4–6 each.
(H and I) Relative expression of CDK inhibitor genes in control (F/+) versus Dpy30 KO (F/−) HSCs in two independent BM chimeras (H) and three FLs at E14.5 (I), as analyzed by RNA sequencing (RNA-seq). The expression levels in F/+ cells were set as 1. The BM HSC data are based on RNA-seq results in donor (Mx1-Cre; Dpy30F/+ or Mx1-Cre; Dpy30F/−)-derived HSCs in the BM chimera recipients in two independent BM transplantations (TP1 and TP2) from our previous work (Yang et al., 2016).
Data are shown as mean ± SD for (C–E). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, by two-tailed Student's t test. See also Figures S1 and S2.