Antibody:	Anti-Perlecan
Detection system:	Biotinylated secondary antibody/[Strept]avidin coupled to alkaline phosphatase
Assay notes:	Fixative: 10% formaldehyde.

Structure	Level	Pattern	Image	Note
TS17: endocardial lining	Present		2A
TS17: pericardium	Present		2A
TS17: primitive ventricle	Present	Regionally restricted	2A	Expression was detected throughout the trabeculated wall of the common ventricular chambers.
TS17: cardiac muscle tissue	Present	Diffuse	2A

Structure	Level	Pattern	Image	Note
TS17: artery	Present		2B	Rostral aorta. At this stage, expression was found dispersed throughout the wall of the major blood vessels. Expression was within the developing smooth muscle cell wall of major blood vessels.

Structure	Level	Pattern	Image	Note
TS17: dorsal aorta	Present		2C	At this stage, expression was found dispersed throughout the wall of the major blood vessels. Expression was within the developing smooth muscle cell wall of major blood vessels.
TS17: future spinal cord	Absent		2C

Structure	Level	Pattern	Image	Note
TS26: arch of aorta	Present		2D	With increasing gestational age, the expression became more localized to the endothelial layer of blood vessels.
TS26: blood vessel endothelium	Present		2D	With increasing gestational age, the expression became more localized to the endothelial layer of blood vessels.

Structure	Level	Pattern	Image	Note
TS21: artery smooth muscle	Present		2E	Expression was within the developing smooth muscle cell wall of major blood vessels.
TS21: dorsal aorta	Present		2E	Expression was within the developing smooth muscle cell wall of major blood vessels.
TS21: lung mesenchyme	Present		2E

Structure	Level	Pattern	Image	Note
TS20: 4th ventricle choroid plexus	Strong		2F
TS20: head mesenchyme	Present	Regionally restricted	2F	Expression was detected in the mesenchyme around the developing blood vessels of the choroid plexus.

Structure	Level	Pattern	Image	Note
TS27: visceral pericardium	Present		2G

Structure	Level	Pattern	Image	Note
TS26: pulmonary artery	Present		2H

Structure	Level	Pattern	Image	Note
TS21: choroid plexus	Present		2I	Expression was localized to the fibrovascular stalk of the choroid plexus.
TS21: brain blood vessel	Present		2I
TS21: cerebral cortex	Absent		2I

Structure	Level	Pattern	Image	Note
TS28: thoracic aorta	Present		2J

Structure	Level	Pattern	Image	Note
TS28: respiratory system blood vessel	Strong		2K	Strong expression was detected in the major pulmonary vessels, both arteries and veins.

Structure	Level	Pattern	Image	Note
TS26: sympathetic ganglion	Present	Regionally restricted	2L	Expression was detected in the mediastinal ganglia surrounding the ventral aorta.

Structure	Level	Pattern	Image	Note
TS20: liver sinusoid	Present		2M	Expression was detected along the sinusoidal lining.

Structure	Level	Pattern	Image	Note
TS25: liver	Present	Regionally restricted	2N	Expression was detected in the portal triad.
TS25: liver sinusoid	Present		2N

Structure	Level	Pattern	Image	Note
TS28: liver parenchyma	Present		2O
TS28: blood vessel endothelium	Present		2O	Expression was primarily expressed in the endothelial layers of the portal and hepatic artery branches.
TS28: hepatic artery	Present		2O	Expression was primarily expressed in the endothelial layers of the portal and hepatic artery branches.
TS28: portal vein	Present		2O	Expression was primarily expressed in the endothelial layers of the portal and hepatic artery branches.

Structure	Level	Pattern	Image	Note
TS21: renal tubule	Present		3A	Expression was detected in the metanephric collecting tubules.
TS21: trunk mesenchyme	Present	Regionally restricted	3A	Expression was detected surrounding the adrenal gland.

Structure	Level	Pattern	Image	Note
TS18: thyroid gland	Present		3B
TS18: 1st arch artery	Present	Regionally restricted	3B	Expression was detected in the mandibular component of the first and third branchial arch artery.
TS18: 3rd arch artery	Present	Regionally restricted	3B	Expression was detected in the mandibular component of the first and third branchial arch artery.

Structure	Level	Pattern	Image	Note
TS20: neurohypophysis infundibulum	Present	Regionally restricted	3C	Expression was detected in the basement membrane zone encasing Rathke's pouch and its adjacent infundibulum.
TS20: remnant of Rathke's pouch	Present	Regionally restricted	3C	Expression was detected in the basement membrane zone encasing Rathke's pouch and its adjacent infundibulum.

Structure	Level	Pattern	Image	Note
TS21: optic stalk	Absent		3D
TS21: eye mesenchyme	Present	Diffuse	3D

Structure	Level	Pattern	Image	Note
TS21: eye mesenchyme	Present	Diffuse	3E

Structure	Level	Pattern	Image	Note
TS26: nerve	Present	Regionally restricted	3F	Several developing myelinated nerve fibers in the facial region showed intense staining.

Structure	Level	Pattern	Image	Note
TS21: dermis	Present	Regionally restricted	3G	Diffuse staining was noted in the upper dermis.
TS21: epidermis	Absent		3G

Structure	Level	Pattern	Image	Note
TS25: head skin	Present	Regionally restricted	3H	In the skin overlying the cartilage of the chondrocranium, expression was localized primarily to discrete regions along the dermo-epidermal zone and around developing hair follicles.
TS25: dermis	Present	Regionally restricted	3H	Expression was detected in the upper dermis around developing hair follicles.

Structure	Level	Pattern	Image	Note
TS28: hindgut	Present	Regionally restricted	3I	Expression was detected in the basement membrane and the mesenchymal component of the hindgut.
TS28: hindgut epithelium	Absent		3I

Structure	Level	Pattern	Image	Note
TS28: stomach	Present	Regionally restricted	3J	Expression was detected in the basement membrane and the mesenchymal component of the stomach.
TS28: stomach epithelium	Absent		3J

Structure	Level	Pattern	Image	Note
TS28: blood vessel	Present	Regionally restricted	3K	Expression was intense in small blood vessels of the cervix and diffuse in the cervical fibromuscular stroma.
TS28: cervix uteri	Present	Regionally restricted	3K	Expression was intense in small blood vessels of the cervix and diffuse in the cervical fibromuscular stroma.
TS28: muscle layer of cervix uteri	Present	Regionally restricted	3K	Expression was intense in small blood vessels of the cervix and diffuse in the cervical fibromuscular stroma.
TS28: epithelial layer of cervix uteri	Absent		3K	The pseudostratified squamous epithelium of the uterine cervix was essentially negative.

Structure	Level	Pattern	Image	Note
TS28: uterus	Present	Regionally restricted		Expression was detected in the capsule and blood vessels.
TS28: mesenchymal stroma of ovary	Present
TS28: vasculature of uterus	Present

Structure	Level	Pattern	Image	Note
TS28: liver	Present	Regionally restricted	3L	Expression was detected in the capsule of the liver.
TS28: arteriole	Present	Regionally restricted	3L	Endothelial staining was noted.

Structure	Level	Pattern	Image	Note
TS28: islets of Langerhans	Present		3M

Structure	Level	Pattern	Image	Note
TS28: epididymis	Present
TS28: tunica albuginea of testis	Present

Structure	Level	Pattern	Image	Note
TS28: adipose tissue	Present			Expression was in fibroadipose tissue.

Structure	Level	Pattern	Image	Note
TS17: vertebral cartilage condensation	Moderate	Diffuse	4A

Structure	Level	Pattern	Image	Note
TS21: vertebral cartilage condensation	Present	Regionally restricted	4B	Expression in cartilage was localized primarily to the hypertrophic zone.

Structure	Level	Pattern	Image	Note
TS22: rib cartilage condensation	Strong		4C

Structure	Level	Pattern	Image	Note
TS22: nucleus pulposus	Strong		4D

Structure	Level	Pattern	Image	Note
TS25: long bone epiphysis	Present	Regionally restricted	4E , 4F	Marked expression was noted in the epiphyseal cartilage of the femur.

Structure	Level	Pattern	Image	Note
TS25: articular system	Present	Regionally restricted	4G	Expression was detected in the articular cartilage.
TS25: cartilage	Present	Regionally restricted	4G	Expression was detected in the articular cartilage.
TS25: femur	Absent		4G	Signal abruptly ended at the adjacent forming femur bone.

Structure	Level	Pattern	Image	Note
TS25: articular system	Present	Regionally restricted	4H	Expression was detected in the articular cartilage.
TS25: cartilage	Present	Regionally restricted	4H	Expression was detected in the articular cartilage.
TS25: clavicle	Absent		4H	Signal abruptly ended at the forming clavicle bone.

Structure	Level	Pattern	Image	Note
TS26: vertebra	Present	Regionally restricted	4I	Expression was most intense in the hypertrophic zones and centers of ossification. The completely ossified region of vertebral bodies was devoid of staining.
TS26: nucleus pulposus	Present		4I

Structure	Level	Pattern	Image	Note
TS27: jaw	Present	Regionally restricted	4J	Expression was detected in the developing cartilage of the jaw.

Structure	Level	Pattern	Image	Note
TS27: vertebra	Present	Regionally restricted	4L	Expression was detected in the vertebral cartilage.