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. 2020 Nov 24;4(22):5810–5824. doi: 10.1182/bloodadvances.2020003299

Figure 1.

Figure 1.

PAR2 deficiency causes a partially penetrant severe embryonic anemia. (A) WT and PAR2−/− embryos were collected between E13.5 and E15.5 from homozygous mating of WT and PAR2−/− parents, and the numbers of normal, pale, and midgestation dead embryos were determined. The number of embryos are shown for individual pregnancies. (B) The numbers of reticulocytes and nRBCs in peripheral blood were counted in 12 normal and 3 pale PAR2−/− embryos at E19.5. Blood smears were stained with Wright solution. Arrows point to nRBC in the blood from 1 of the pale embryos. (C) PAR2+/− embryos were collected at E13.5 from mating of WT or PAR2−/− mother with PAR2−/− or WT father, respectively, and the numbers of normal, pale, and dead embryos, as well as embryo and placenta weight, were determined. Data are mean ± standard deviation (SD). **P < .01, unpaired Student t test; not significant (ns), Mann-Whitney U test. (D) PAR2+/− and PAR2−/− embryos were collected at E13.5 from mating of PAR2+/− mother with PAR2−/− father, and the numbers of normal, pale, and dead embryos, as well as embryo weights, were determined. Data are mean ± SD. ns, unpaired Student t test. (E) We used an independently targeted line to collect PAR2+/− and PAR2−/− embryos at E13.5 to E17.5 from the mating of a PAR2+/− mother with a PAR2−/− father. Examples of PAR2+/− and PAR2−/− embryos with pale (red boxes) or midgestation lethal phenotypes shown. (F) Livers from WT adult mice, as well as fetal livers from WT embryos at E13.5 and E15.5, were collected, and the mRNA expression of coagulation factors prothrombin, FVII, and FX was normalized to r18s as a housekeeping gene. The percentage above the columns shows the relative expression of coagulation factors at each embryonic stage in comparison with adult liver measured in parallel. ****P < .0001, 1-way ANOVA with Dunnett multiple-comparisons test.