Fig. 3

notch3^fh332 mutant larvae have brain hemorrhage and a defective BBB. (A-D) Examples of brain hemorrhage in 3 dpf notch3^fh332 mutant larvae. (A-C) Lateral view. (D) Dorsal view. Arrowheads indicate blood pooling in the brain ventricle. Insets show high magnification view of the hemorrhage in the brain ventricle. (E-H) Representative results of BBB assay in 4 dpf wild-type (WT) and mutant larvae. (E,F) Lateral views of the brain. (G,H) Dorsal views of the midbrain and hindbrain. Mutant embryos failed to retain 2000 kD Alexa488-dextran in the brain microvessels 90 minutes after the injection. Arrowheads and arrows indicate brain vessels in the wild-type and mutant larvae, respectively. (I-L) Immunostaining of 3 dpf wild-type and mutant larvae with Claudin 5 and ZO-1 antibodies. Arrowheads indicate Claudin 5 or ZO-1 staining at the blood vessels. Open arrowheads indicate ZO-1 staining along brain ventricular wall. Scale bar: 200 μm.