Cerebral hemorrhage in Nfe2^−/− mice occurs as early as E11.5 and persists through development. (A) Representative images of Nfe2^−/− and age-matched Nfe2^+/+ control embryos at E11.5 (Nfe2^+/+ n = 16; Nfe2^−/− n = 25), E12.5 (Nfe2^+/+ n = 13; Nfe2^−/− n = 18), E13.5 (Nfe2^+/+ n = 16; Nfe2^−/− n = 14), and E14.5 (Nfe2^+/+ n = 14; Nfe2^−/− n = 14). Scale bars, 1 mm. (B-C) Scored bleeding phenotypes in the Nfe2^+/+ and Nfe2^−/− embryo body (B) or head (C) from E10.5 to E14.5. (D) Bleeding into the lymphatic sacs and lymphatic vessels is seen at E14.5 in the Nfe2^−/− but not in the Nfe2^+/+ mice. Immunofluorescence staining of E14.5 Nfe2^+/+ skin showing that erythrocytes (TER119, white) are restricted to the blood vessels (PECAM1, red), whereas erythrocytes are readily detected within dilated lymphatic vessels (LYVE1, green) in Nfe2^−/− mice. Embryo scale bars, 2 mm. Skin confocal scale bars, 100 µm. (E-F). Frequency of bleed types at E14.5 (Nfe2^+/+ n = 14; Nfe2^−/− n = 12) (E) and E18.5 in Nfe2^+/+ (n = 10) and Nfe2^−/− embryos (n = 10) (F). Scale bars for whole embryos, 1 mm; scale bars for skin, 150 μm. Contingency table analysis was performed using the 2-tailed χ² test for the comparison of bleed frequencies (B-C). *P < .05; ****P < .0001. ns, not significant.