Extended Data Fig. 8: Characterization of Evi1+/− embryos.
From: Independent origins of fetal liver haematopoietic stem and progenitor cells
a, Analysis of E12.5 Evi1+/− embryos. Left, Representative images of E12.5 embryos. Scale bars, 200 μm. Right, Quantitation of c-Kit+ cells in the fetal liver (Evi1+/+ mice, n = 5 ; Evi1+/− mice, n = 7). b, Normal hematopoietic cluster formation in Evi1+/− embryos. Left, Whole-mount immunostaining of Evi1+/+ (34 sp) and Evi1+/− (33 sp) embryos for c-Kit (green) and CD31 (magenta) expression. Scale bars, 100 μm. Right, Number of c-Kit+ cells localized in the middle segment of the dorsal aorta (DA). Middle segment is 7 somite-lengths18. Evi1+/+ (n = 3, 33–35 sp). Evi1+/− (n = 3, 33 and 34 sp). c, Specific decrease in stem cell fraction in Evi1 heterozygous embryos at E14.5. Left, Representative flow cytometry plots. Right, Frequency of HSC and progenitor fractions (Evi1+/+ mice, n = 8 ; Evi1+/− mice, n = 6). Similar to the results at the E12.5 stage (Fig. 3b), severe defects were observed in the HSC fractions (15-fold decrease) in Evi1+/− embryos at this time point. d, Kinetic analysis of HSC and progenitor formation from HlfCreERT2-labeled cells in Evi1+/− embryos. Top left, Schematic of tamoxifen treatment and analysis. Bottom left, Representative flow cytometry plots of HlfCreERT2-labeled cells (red dots). Right, Quantitation of tdTomato+ cells in the fetal liver (E11.5 Evi1+/+, n = 6; E12.5, Evi1+/+ n = 5; E11.5, Evi1+/− n = 5; E12.5 Evi1+/−, n = 5). All error bars represent means ± SD. Statistical analysis was performed two-sided unpaired Student’s t-test (a–c).
