Antibody:	anti-cleaved Notch1 (Val1744) antibody (anti-NICD) (#2421)
Detection system:	Biotinylated secondary antibody/[Strept]avidin coupled to Cy3
Assay notes:	Antibody detects cytosolic domain of Notch1 only when cleaved between Gly1743 and Val1744.

Structure	Level	Pattern	Image	Note
TS10: body cavity or lining	Present		1E , 1D	Expressed in the intraembryonic mesoderm region.
TS10: embryo ectoderm	Absent		1D , 1E

Structure	Level	Pattern	Image	Note
TS11: allantois	Present	Regionally restricted	1F , 1G	Expressed in developing vessels.
TS11: cardiogenic plate	Strong	Regionally restricted	1G , 1H	Expressed in cardiac crescent cells, primitive endocardium.
TS11: amnion	Strong		1G , 1H
TS11: head mesenchyme	Present	Single cells	1H , 1G	Expressed in a few cells.

Structure	Level	Pattern	Image	Note
TS11: blood island	Present		1I

Structure	Level	Pattern	Image	Note
TS12: head mesenchyme	Present	Scattered	1J
TS12: dorsal aorta	Present		1J , 1K
TS12: endocardial tube	Present		1J , 1K
TS12: amnion	Present		1J

Structure	Level	Pattern	Image	Note
TS13: primitive ventricle endocardial tube	Present	Regionally restricted	2B , 2A	Expression was restricted to endocardial cells adjacent to the myocardium, while distal endocardial cells expressed low levels.

Structure	Level	Pattern	Image	Note
TS14: atrioventricular canal	Present	Regionally restricted	2E , 2D	Expressed in endocardial cells. Expression in prevalve territories was highest.

Structure	Level	Pattern	Image	Note
TS15: outflow tract endocardial tube	Present		2G , 2F

Structure	Level	Pattern	Image	Note
TS15: atrioventricular canal endocardial lining	Present		2J , 2K

Structure	Level	Pattern	Image	Note
TS17: outflow tract endocardial tube	Present		2O , 2N

Structure	Level	Pattern	Image	Note
TS17: atrioventricular canal endocardial lining	Present		2P , 2Q
TS17: atrioventricular endocardial cushion	Present	Regionally restricted	2Q	Expressed in the endocardial cells. No expression in mesenchymal cells.

Structure	Level	Pattern	Image	Note
TS15: atrium endocardial lining	Present	Widespread	2R
TS15: primitive ventricle endocardial lining	Present	Non-Uniform	2S , 2T , 2R	Expressed unequally in the developing trabeculae. Stronger in endocardium at the base of trabeculae and weaker in distal endocardium.

Structure	Level	Pattern	Image	Note
TS19: outflow tract endothelium	Strong		3C , 3B

Structure	Level	Pattern	Image	Note
TS19: atrioventricular canal endocardial lining	Strong		3E , 3D
TS19: left ventricle endocardial lining	Strong		3D , 3F	Trabecular endocardium.
TS19: right ventricle endocardial lining	Strong		3D , 3G	Trabecular endocardium.

Structure	Level	Pattern	Image	Note
TS21: semilunar valve	Present	Regionally restricted	3H , 3I	Expressed in the endocardium.
TS21: heart endothelium	Present	Regionally restricted	3H , 3I	Decreased expression in arterial endothelium.
TS21: artery smooth muscle	Present	Regionally restricted	3I	Detected in smooth muscle cells of the heart arterial wall.
TS21: left atrium endocardial lining	Present	Regionally restricted	3J , 3K	Expressed in endocardium lining the developing trabeculae.

Structure	Level	Pattern	Image	Note
TS21: mitral valve	Present	Regionally restricted	3M , 3L	Expressed in the endocardium but staining is reduced in the valve component facing the atrium.
TS21: right ventricle endocardial lining	Strong		3N , 3L
TS21: coronary artery	Strong	Regionally restricted	3O , 3L	Expressed in the developing coronary vessels of the left ventricle.

Structure	Level	Pattern	Image	Note
TS22: semilunar valve	Present	Regionally restricted	3Q	Expressed in the endocardium, but not in the arterial endothelium.
TS22: outflow tract endothelium	Present	Regionally restricted	3Q	Expressed in the endocardium, but not in the arterial endothelium.

Structure	Level	Pattern	Image	Note
TS22: tricuspid valve	Present	Regionally restricted	3S	Expressed in the endocardial lining.

Structure	Level	Pattern	Image	Note
TS13: dorsal aorta	Strong	Regionally restricted	4A , 4B	Expressed in endothelial cells.
TS13: primary head vein	Absent		4B , 4A

Structure	Level	Pattern	Image	Note
TS13: arterial blood vessel	Present	Regionally restricted	4D	Expressed in the endothelium connecting the dorsal aorta with the second arterial arch.

Structure	Level	Pattern	Image	Note
TS15: dorsal aorta	Present		4E
TS15: anterior cardinal vein	Absent		4E

Structure	Level	Pattern	Image	Note
TS15: dorsal aorta	Present	Regionally restricted	4M	Expressed in endothelial cells.
TS15: mesonephros mesenchyme	Present	Regionally restricted	4M	Expressed in the urogenital ridge.

Structure	Level	Pattern	Image	Note
TS17: dorsal aorta	Present	Regionally restricted	4N	Expressed in endothelial cells.

Structure	Level	Pattern	Image	Note
TS15: dorsal aorta	Present	Regionally restricted	4O	Expressed in hematopoietic cluster cells closer to the aorta endothelium, but not in cells budding from the aorta.

Structure	Level	Pattern	Image	Note
TS17: dorsal aorta	Present	Regionally restricted	4P	Expressed in hematopoietic cluster cells closer to the aorta endothelium, but not in cells budding from the aorta.

Structure	Level	Pattern	Image	Note
TS19: dorsal aorta	Present	Regionally restricted	4Q	Expression in arterial endothelium begins to disappear, while smooth muscle cells are stained.
TS19: cardiovascular system endothelium	Present	Regionally restricted	4Q	Expression in arterial endothelium begins to disappear, while still no expression in endothelium of anterior cardinal vein.

Structure	Level	Pattern	Image	Note
TS21: dorsal aorta	Present	Regionally restricted	4R	Expressed in smooth muscle cells. No expression in endothelial cells.

Structure	Level	Pattern	Image	Note
TS15: hepatic primordium	Present	Regionally restricted	4U	Expressed in the vasculature.

Structure	Level	Pattern	Image	Note
TS17: hepatic primordium	Present	Regionally restricted	4V	Expressed in the vasculature.

Structure	Level	Pattern	Image	Note
TS21: meninges	Present	Regionally restricted	4X	Expressed in the vasculature of the meningeal plexus.

Structure	Level	Pattern	Image	Note
TS21: lung vascular element	Present	Regionally restricted	4Y	Expressed in arteries.

Structure	Level	Pattern	Image	Note
TS13: trunk unsegmented mesenchyme	Strong		5A , 5B	Presomitic mesoderm.

Structure	Level	Pattern	Image	Note
TS15: somite	Absent		5D , 5E

Structure	Level	Pattern	Image	Note
TS15: tail unsegmented mesenchyme	Present	Regionally restricted	5G	Expressed in the cleft-forming area between S-1 and S0 and the posterior presomitic mesoderm.

Structure	Level	Pattern	Image	Note
TS15: nephrogenic cord	Present		5H
TS15: nephric duct	Absent		5H

Structure	Level	Pattern	Image	Note
TS15: nephrogenic cord	Present		5I
TS15: nephric duct	Absent		5I

Structure	Level	Pattern	Image	Note
TS17: mesonephric tubule	Present		5K

Structure	Level	Pattern	Image	Note
TS21: renal vesicle	Present		5L , 5M
TS21: mesonephric tubule	Absent		5L , 5M
TS21: nephric duct, mesonephric portion	Absent		5L , 5M

Structure	Level	Pattern	Image	Note
TS22: comma-shaped body	Present		5N
TS22: s-shaped body	Present		5N

Structure	Level	Pattern	Image	Note
TS21: hair follicle	Present		5O
TS21: vibrissa follicle	Present		5O

Structure	Level	Pattern	Image	Note
TS22: ear epithelium	Present		5P

Structure	Level	Pattern	Image	Note
TS21: esophagus epithelium	Present		5Q

Structure	Level	Pattern	Image	Note
TS21: pancreas	Present	Regionally restricted	5R , 5S	Expressed in the pancreatic artery and in the acini.
TS21: pancreatic acinus	Present		5S , 5R

Structure	Level	Pattern	Image	Note
TS21: thymus primordium	Present	Regionally restricted	5U , 5T	Expressed in developing thymocytes and stromal cells.

Structure	Level	Pattern	Image	Note
TS22: thymus primordium	Strong		5V

Structure	Level	Pattern	Image	Note
TS15: future hindbrain	Present	Widespread	6A	Expressed throughout the hindbrain, particularly in the distal or abluminal region, and with less intensity in cells closest to the luminal side of the neural canal.

Structure	Level	Pattern	Image	Note
TS17: forebrain	Present	Widespread	6F , 6E	Expressed throughout the neuroepithelium.

Structure	Level	Pattern	Image	Note
TS17: hindbrain	Present	Widespread	6G	Expressed throughout the neuroepithelium.

Structure	Level	Pattern	Image	Note
TS19: telencephalon	Present	Regionally restricted	6I , 6J	Expressed at the roof, in nuclei close to the ventricular zone.
TS19: diencephalon lateral wall	Present		6I	Expressed in nuclei close to the ventricular zone.
TS19: diencephalon roof plate	Present		6I	Expressed in nuclei close to the ventricular zone.

Structure	Level	Pattern	Image	Note
TS19: dorsal root ganglion	Present		6K
TS19: neural tube ventricular layer	Present		6K
TS19: neural tube lateral wall	Present	Regionally restricted	6K	Expressed in cells of the ventricular zone. No expression in postmitotic ventral horn cells.

Structure	Level	Pattern	Image	Note
TS21: diencephalon	Present	Regionally restricted	6N , 6M	Expressed in cells near the neural canal.

Structure	Level	Pattern	Image	Note
TS21: spinal cord ventricular layer	Present		6O	The pattern was similar to BrdU staining.

Structure	Level	Pattern	Image	Note
TS17: hindbrain ventricular layer	Strong		7B , 7A
TS17: trigeminal ganglion	Present		7A , 7B
TS17: primary head vein	Absent		7A , 7B

Structure	Level	Pattern	Image	Note
TS19: trigeminal ganglion	Present		7C , 7D

Structure	Level	Pattern	Image	Note
TS21: spinal cord ventricular layer	Strong		7E
TS21: dorsal root ganglion	Strong		7E , 7F	Expressing cells and nonexpressing cells are mixed.

Structure	Level	Pattern	Image	Note
TS14: otic pit	Present		7G , 7H

Structure	Level	Pattern	Image	Note
TS17: otocyst	Present	Regionally restricted	7I , 7J	Expressed in the ventral lateral region.
TS17: acoustic ganglion	Present		7I , 7J

Structure	Level	Pattern	Image	Note
TS19: endolymphatic appendage	Present		7L , 7K
TS19: semicircular canal	Absent		7L , 7K

Structure	Level	Pattern	Image	Note
TS21: cochlea epithelium	Present		7M
TS21: semicircular canal epithelium	Present		7M

Structure	Level	Pattern	Image	Note
TS21: cochlea epithelium	Present		7N
TS21: semicircular canal epithelium	Present		7N

Structure	Level	Pattern	Image	Note
TS22: cochlear duct epithelium	Present	Regionally restricted	7P , 7O	Expressed in the mid-basal region.

Structure	Level	Pattern	Image	Note
TS19: ophthalmic artery	Present	Regionally restricted	7Q	Expressed in the hyaloid artery and vasculature around the eye.
TS19: neural retina	Weak	Regionally restricted	7Q	Very weakly in the peripheral region.
TS19: optic stalk	Present		7Q

Structure	Level	Pattern	Image	Note
TS21: neural retina	Present	Regionally restricted	7R	Increased expression, but was excluded from the central region.
TS21: ophthalmic artery	Present	Regionally restricted	7R	Expressed in the hyaloid plexus.

Structure	Level	Pattern	Image	Note
TS22: neural retina	Present	Regionally restricted	7S	Increased expression, but was excluded from the central region.
TS22: ophthalmic artery	Present	Regionally restricted	7S	Expressed in the hyaloid plexus.

Structure	Level	Pattern	Image	Note
TS19: nasal epithelium	Present	Regionally restricted	7U , 7V	Expressed in the basal layer.
TS19: nasal mesenchyme	Present	Regionally restricted	7V , 7U	Expressed in the vasculature surrounding the basal layer.

Structure	Level	Pattern	Image	Note
TS21: nasal cavity epithelium	Present	Regionally restricted	7W , 7X	Expressed in the epithelium of the primitive nasal cavity and serous gland.

Structure	Level	Pattern	Image	Note
TS15: neural tube	Present		8A , 8B	Strongly reduced expression compared with wild-type.
TS15: outflow tract endocardial tube	Present		8A , 8C	Strongly reduced expression compared with wild-type.
TS15: primitive ventricle endocardial lining	Present		8A , 8C	Strongly reduced expression compared with wild-type.

Structure	Level	Pattern	Image	Note
TS15: neural tube	Present		8D , 8E	Strongly reduced expression compared with wild-type.
TS15: outflow tract endocardial tube	Present		8F , 8D	Strongly reduced expression compared with wild-type.
TS15: primitive ventricle endocardial lining	Present		8F , 8D	Strongly reduced expression compared with wild-type.

Structure	Level	Pattern	Image	Note
TS15: paraxial mesenchyme	Present		8H , 8G	Strongly reduced expression compared with wild-type.

Structure	Level	Pattern	Image	Note
TS15: neural tube	Absent		S2B , S2A	Only weak and dispersed background non-nuclear staining was observed.
TS15: heart	Absent		S2C , S2A	Only weak and dispersed background non-nuclear staining was observed.

Structure	Level	Pattern	Image	Note
TS15: atrioventricular canal endocardial lining	Present		S7

Structure	Level	Pattern	Image	Note
TS15: primitive ventricle endocardial lining	Present	Non-Uniform	S8	Expressed unequally in the developing trabeculae. Stronger in endocardium at the base of trabeculae and weaker in distal endocardium.

Structure	Level	Pattern	Image	Note
TS21: coronary artery	Strong		S9B

Structure	Level	Pattern	Image	Note
TS21: coronary artery	Strong		S9D

Structure	Level	Pattern	Image	Note
TS21: coronary artery	Strong		S9F

Structure	Level	Pattern	Image	Note
TS12: future hindbrain neural fold	Present	Scattered		Expressed in the dorsal cells.
TS12: future midbrain neural fold	Present	Scattered		Expressed in the dorsal cells.