Probe:	Bigh3 probe1
Probe preparation:	Antisense labelled with S35 DNA
Visualized with:	Autoradiography
Assay notes:	Two antisense oligo probes were described, but the one used in this assay was not specified. Either MGI:6273193 or MGI:6273196 could have been used.

Structure	Level	Pattern	Image	Note
TS20: central nervous system	Absent
TS20: dermis	Absent
TS20: heart	Absent
TS20: liver	Absent
TS20: skin	Absent
TS20: embryo mesenchyme	Present	Regionally restricted		Expression was detected in the mesenchyme of the first and second branchial arches, but not of the third.
TS20: frontal bone primordium	Present
TS20: head bone	Present	Regionally restricted		Expression was detected in the frontal, parietal, sphenoid, and temporal bones.
TS20: head mesenchyme derived from neural crest	Present	Regionally restricted		Expression was detected in the neural crest derived mesenchyme of the VIII, IX, and X cranial nerve nuclei.

Structure	Level	Pattern	Image	Note
TS19: nasal pit	Absent
TS19: optic stalk	Absent
TS19: eye mesenchyme	Present	Regionally restricted		Expression was detected in the mesenchyme surrounding the optic stalk.
TS19: rib pre-cartilage condensation	Present	Regionally restricted		Expression was detected in the costal and vertebral mesenchyme.
TS19: vertebral pre-cartilage condensation	Present	Regionally restricted		Expression was detected in the costal and vertebral mesenchyme.
TS19: cardiovascular system mesenchyme	Present	Regionally restricted		Expression was detected in mesenchyme surrounding the developing vessels.
TS19: blood vessel endothelium	Absent

Structure	Level	Pattern	Image	Note
TS20: ascending aorta	Present
TS20: dorsal root ganglion	Present
TS20: fronto-nasal process	Present
TS20: inferior atrioventricular endocardial cushion	Present
TS20: pulmonary artery	Present
TS20: superior atrioventricular endocardial cushion	Present
TS20: lung	Present	Regionally restricted		Expression was exclusively peribronchiolar labeling. No staining was detected in the bronchiolar and lung epithelia.
TS20: lung epithelium	Absent
TS20: remnant of Rathke's pouch	Absent
TS20: tongue muscle	Absent
TS20: trunk mesenchyme	Present	Regionally restricted		The myotome-derived premuscular mass (lateral mesoderm) was labeled.
TS20: head mesenchyme	Present	Regionally restricted		The mesenchyme surrounding Rathke's pouch is labeled.
TS20: head bone	Present	Regionally restricted		Expression was detected in the ethmoidis.
TS20: axial skeleton	Present	Regionally restricted		Expression was detected in the intervertebral disks.
TS20: oral region mesenchyme	Present	Regionally restricted		Vascular structures under the future tongue are labeled in the subendothelial mesenchyme.
TS20: thymus primordium	Present
TS20: gut	Absent			The pharyngeal gut and all the upper gut as well as the mid- and hindgut are not labeled.
TS20: nephric duct	Absent
TS20: chondrocranium bone	Present	Regionally restricted		Expression was detected in the basioccipital and chondro occiput.
TS20: bone	Present	Regionally restricted		Expression was detected in the anterior thoracic bone wall (sternum and ribs primordia).
TS20: heart	Present	Regionally restricted		Expression was detected in the mesenchyme of the ombilical cordis.

Structure	Level	Pattern	Image	Note
TS20: heart	Present		1A/B

Structure	Level	Pattern	Image	Note
TS21: Meckel's cartilage	Present
TS21: ethmoid bone primordium	Present
TS21: nasal capsule	Present
TS21: cranium	Present	Regionally restricted		Expression was detected in the base of the skull.
TS21: dorsal root ganglion	Present
TS21: thoracic body wall	Present	Regionally restricted		The whole anterior thoracic wall was labeled.
TS21: pancreas	Present
TS21: adrenal gland	Present	Regionally restricted		Expression was detected in the capsule of the adrenal gland, but the cortex and medulla were not labeled.
TS21: gut muscularis	Present
TS21: gut epithelium	Absent

Structure	Level	Pattern	Image	Note
TS21: lung	Strong	Regionally restricted	1C/D	Strong expression was detected in the mesenchyme surrounding the bronchioles, but there was no label in the bronchiolar epithelium.
TS21: lung mesenchyme	Strong	Regionally restricted	1C/D	Strong expression was detected in the mesenchyme surrounding the bronchioles, but there was no label in the bronchiolar epithelium.

Structure	Level	Pattern	Image	Note
TS21: trunk mesenchyme	Present	Regionally restricted	1E/F	Expression was observed in the mesenchyme surrounding the kidney.
TS21: mesenchyme of metanephros	Present	Regionally restricted	1E/F	Expression was detected in the periglomerular and tubular mesenchyme.
TS21: capillary loop renal corpuscle	Absent		1E/F
TS21: renal tubule	Absent		1E/F

Structure	Level	Pattern	Image	Note
TS22: central nervous system	Absent
TS22: leptomeninges	Present
TS22: peritoneum	Present
TS22: vault of skull	Present
TS22: gut	Present			The labeling of the kidney, lung, and gut was unchanged from E13.5.
TS22: lung	Present			The labeling of the kidney, lung, and gut was unchanged from E13.5.
TS22: metanephros	Present			The labeling of the kidney, lung, and gut was unchanged from E13.5.
TS22: inner ear mesenchyme	Present	Regionally restricted		Expression was detected in the mesenchyme surrounding the developing bones of the inner ear.

Structure	Level	Pattern	Image	Note
TS22: perichondrium	Strong		1G/H

Structure	Level	Pattern	Image	Note
TS21: central nervous system	Absent		2A/D
TS21: skin	Absent		2A/D
TS21: rib cartilage condensation	Present		2A/D
TS21: rib pre-cartilage condensation	Present		2A/D
TS21: vertebral cartilage condensation	Present		2A/D
TS21: vertebral pre-cartilage condensation	Present		2A/D

Structure	Level	Pattern	Image	Note
TS22: cartilage	Present	Regionally restricted	2B/E	Expression was detected in active zones of growth.
TS22: limb cartilage condensation	Present	Regionally restricted	2B/E	Expression was detected in active zones of growth.

Structure	Level	Pattern	Image	Note
TS25: cornea	Present		2C/F
TS25: cornea epithelium	Present		2C/F
TS25: cornea stroma	Present		2C/F
TS25: optic choroid	Present		2C/F
TS25: sclera	Present		2C/F
TS25: lens	Absent		2C/F
TS25: neural retina	Absent		2C/F
TS25: pigmented retinal epithelium	Absent		2C/F

Structure	Level	Pattern	Image	Note
TS21: choroid plexus	Present		3A/B

Structure	Level	Pattern	Image	Note
TS22: rib cartilage condensation	Weak		3C/D
TS22: pectoralis	Absent		3C/D
TS22: trunk connective tissue	Present	Regionally restricted	3C/D	Expression was detected in the capsule of Glisson.

Structure	Level	Pattern	Image	Note
TS22: eye	Present	Regionally restricted	3E	Posterior labeling of the sclera and choroid with transcription progressing toward the thickness of the connective tissue of the future eyelids.
TS22: eyelid	Present	Regionally restricted	3E	Posterior labeling of the sclera and choroid with transcription progressing toward the thickness of the connective tissue of the future eyelids.

Structure	Level	Pattern	Image	Note
TS22: eye	Present	Regionally restricted	3F	Posterior labeling of the sclera and choroid with transcription progressing toward the thickness of the connective tissue of the future eyelids.
TS22: eyelid	Present	Regionally restricted	3F	Posterior labeling of the sclera and choroid with transcription progressing toward the thickness of the connective tissue of the future eyelids.

Structure	Level	Pattern	Image	Note
TS23: midgut muscularis	Present	Regionally restricted	3G/H	Expression was detected in the smooth muscles surrounding the midgut.

Structure	Level	Pattern	Image	Note
TS23: perioptic mesenchyme	Present	Regionally restricted		The periocular mesenchyme posterior to the lens was labeled.
TS23: cornea	Absent
TS23: optic choroid	Present
TS23: choroid plexus	Present
TS23: dermis	Present
TS23: frontal bone primordium	Present
TS23: intervertebral disc	Present
TS23: leptomeninges	Present
TS23: parietal bone	Present
TS23: pericardium	Present
TS23: peritoneum	Present
TS23: connective tissue	Present	Regionally restricted		Expression was detected in the connective tissue located beneath the olfactive epithelium. Expression was also noted in the capsule of Glisson.
TS23: esophagus muscularis	Present	Regionally restricted		Expression was detected in the smooth muscle surrounding the esophagus and other parts of the gut.
TS23: gut muscularis	Present	Regionally restricted		Expression was detected in the smooth muscle surrounding the esophagus and other parts of the gut.
TS23: trunk mesenchyme	Present	Regionally restricted		Expression was detected in the mesenchyme surrounding the ribs and vertebrae.
TS23: lung mesenchyme	Present	Regionally restricted		Expression was detected in the mesenchyme surrounding the bronchioles.
TS23: mesenchyme of metanephros	Present	Regionally restricted		Expression was detected in the mesenchyme surrounding the kidney tubules.
TS23: bone	Present	Diffuse
TS23: central nervous system	Absent

Structure	Level	Pattern	Image	Note
TS24: face mesenchyme	Present	Regionally restricted		Expression was detected in the mesenchyme surrounding the vibrissae.
TS24: intervertebral disc	Present
TS24: lung	Present
TS24: metanephros	Present
TS24: nucleus pulposus	Present
TS24: perichondrium	Present
TS24: secondary palate	Present
TS24: stomach muscularis externa	Present
TS24: submandibular gland mesenchyme	Present
TS24: thymus	Present
TS24: tooth	Present
TS24: jaw bone	Present	Regionally restricted		Expression was detected in the developing membranous dental bone.
TS24: connective tissue	Present	Regionally restricted		Expression was detected in the perineurium of the dental nerve, as well as the connective tissue surrounding the submaxillary acini, pancreatic acini, and the Langerhans islands of the pancreas.
TS24: central nervous system	Absent
TS24: liver	Absent
TS24: stomach epithelium	Absent

Structure	Level	Pattern	Image	Note
TS25: esophagus muscularis	Present
TS25: gut muscularis	Present
TS25: trachea smooth muscle	Present
TS25: perichondrium	Strong	Regionally restricted		Expression was strong in active zones of bone growth.
TS25: dermis connective tissue	Absent
TS25: cardiac muscle tissue	Present
TS25: pericardium	Present
TS25: peritoneum	Present
TS25: pleura	Present
TS25: tongue muscle	Present
TS25: alveolar system	Present	Regionally restricted		The mesenchymal component of the developing alveolocapillary membrane was labeled.
TS25: connective tissue	Present	Regionally restricted		The capsule of Glisson was labeled.
TS25: coronary artery	Present
TS25: heart valve	Present
TS25: central nervous system	Absent
TS25: epidermis	Absent
TS25: liver	Absent

Structure	Level	Pattern	Image	Note
TS26: cornea epithelium	Present			Expression was mostly confined to the anterior part of the cornea where the epithelium appeared more labeled than the stroma.
TS26: cornea stroma	Present			Expression was mostly confined to the anterior part of the cornea where the epithelium appeared more labeled than the stroma.
TS26: retina	Absent
TS26: ligament	Present	Regionally restricted		Intercostal ligaments were labeled.
TS26: axial skeleton	Present	Regionally restricted		Perivertebral and pericostal regions were labeled.
TS26: liver	Absent
TS26: lung	Absent
TS26: metanephros	Absent
TS26: choroid plexus	Present
TS26: colon	Present