Antibody:	Anti-C21orf63
Detection system:	Secondary antibody coupled to Alexa Fluor 594

Structure	Level	Pattern	Image	Note
TS17: dorsal root ganglion	Present		1A
TS17: future spinal cord	Present	Regionally restricted	1C , 1A , 1B	Expression was detected in motor neurons in the motor column and by their axons in the motor root. Immunopositive sensory axons were noted entering the dorsal horn. A small number of commissural axons were seen passing along the inside border of the motor column and then across the midline floor plate.

Structure	Level	Pattern	Image	Note
TS25: spinal cord	Present	Regionally restricted	1F , 1E , 1D	Expression was detected in the ascending and descending longitudinal pathways that consititute the dorsal columns, dorsal funiculus, and ventrolateral funiculus. There was no expression in the motor column, spinal motor axons, the dorsal root entry zone, or commissural axons as they coursed towards the midline and crossed the floor plate.
TS25: dorsal root ganglion	Absent		1D

Structure	Level	Pattern	Image	Note
TS22: main olfactory bulb external plexiform layer	Strong		2A
TS22: main olfactory bulb olfactory nerve layer	Strong		2A
TS22: olfactory nerve	Strong		2A
TS22: main olfactory bulb layer	Strong	Regionally restricted	2A	Strong expression was detected throughout the outer layers of the olfactory bulb, which include the olfactory sensory axons.

Structure	Level	Pattern	Image	Note
TS25: main olfactory bulb external plexiform layer	Strong		2B , 2E , 2H
TS25: main olfactory bulb olfactory nerve layer	Strong		2B , 2E , 2H
TS25: main olfactory bulb layer	Strong	Regionally restricted	2E , 2H , 2B	Strong expression was detected throughout the outer layers of the olfactory bulb, which include the olfactory sensory axons (as indicated by co-expression with Gap43).

Structure	Level	Pattern	Image	Note
TS27: main olfactory bulb external plexiform layer	Strong		2C
TS27: main olfactory bulb olfactory nerve layer	Strong		2C
TS27: main olfactory bulb layer	Strong	Regionally restricted	2C	Strong expression was detected throughout the outer layers of the olfactory bulb, which include the olfactory sensory axons.

Structure	Level	Pattern	Image	Note
TS28: main olfactory bulb olfactory nerve layer	Strong		2I , 2L , 2D
TS28: main olfactory bulb layer	Strong	Regionally restricted	2L , 2I , 2D	Strong expression was detected throughout the outer layers of the olfactory bulb, which include the olfactory sensory axons (as indicated by co-expression with Gap43).
TS28: main olfactory bulb glomerular layer	Weak	Regionally restricted	2L , 2D , 2I	Expression was detected exclusively in the pre-terminal portions of olfactory sensory axons within the nerve fiber layer and as the axons enter the glomerular layer, but it is not present in the glomerular neuropil containing the terminal portions of the sensory axons.
TS28: main olfactory bulb external plexiform layer	Strong	Punctate	2L , 2D , 2I

Structure	Level	Pattern	Image	Note
TS22: neural retina	Present	Regionally restricted	3A , 3D	Expression was high in retinal ganglion cells and their axons in the inner most layer of the retina.
TS22: optic nerve	Present		3A , 3D

Structure	Level	Pattern	Image	Note
TS25: retina layer	Present		3E , 3H	Expression was detected in the retinal ganglion cells and their axons in the inner most layer of the retina.
TS25: optic nerve	Present		3E , 3H
TS25: retina nerve fiber layer	Present		3E , 3H

Structure	Level	Pattern	Image	Note
TS22: optic chiasma	Present		3I

Structure	Level	Pattern	Image	Note
TS25: suprachiasmatic nucleus	Weak	Scattered	3J	A weak mosaic staining pattern was noted.
TS25: optic chiasma	Present		3J

Structure	Level	Pattern	Image	Note
TS27: suprachiasmatic nucleus	Present	Widespread	3K
TS27: optic chiasma	Present		3K

Structure	Level	Pattern	Image	Note
TS28: suprachiasmatic nucleus	Trace	Scattered	3L	Expression decreased to pre-E17.5 levels by P10.
TS28: optic chiasma	Present		3L

Structure	Level	Pattern	Image	Note
TS22: cortical plate	Present	Regionally restricted	4A	Expression was detected in the cortical plate of the cingulate cortex.
TS22: neocortex	Present	Regionally restricted	4A	Expression was detected in the cortical plate of the cingulate cortex, but not in the underlying deeper layers of the cingulate cortex, including the intermediate zone and germinal layer.
TS22: fornix	Present	Regionally restricted	4A	Expression was seen in axons projecting into the fornix.

Structure	Level	Pattern	Image	Note
TS25: cerebral cortex intermediate zone	Strong	Regionally restricted	4B	Expression was strong in the intermediate zone underlying the cingulate cortex and more weakly expressed in the axons from the intermediate zone of the remaining neocortex.
TS25: corpus callosum	Weak	Regionally restricted	4H , 4E , 4B	At the midline, the cingulate axons downregulate expression and diffusely course in the dorsal corpus callosum. Neocortical axons continue weak expression as crossing the midline in the ventral corpus callosum.
TS25: brain commissure	Strong	Regionally restricted	4B	Robust expression was noted in the hippocampal commissure.

Structure	Level	Pattern	Image	Note
TS27: cerebral cortex intermediate zone	Strong	Regionally restricted	4C	Strong expression was detected in the intermediate zone of the cingulate cortex and neocortex.
TS27: cingulate cortex	Strong	Regionally restricted	4C	Strong expression was detected in the intermediate zone of the cingulate cortex and neocortex.
TS27: neocortex	Strong	Regionally restricted	4C	Strong expression was detected in the intermediate zone of the cingulate cortex and neocortex.
TS27: corpus callosum	Strong	Regionally restricted	4L , 4I , 4C	Strong co-expression with Gap43-positive commissural axons through the dorsoventral depth of the corpus callosum.
TS27: hippocampal commissure	Present		4C

Structure	Level	Pattern	Image	Note
TS28: corpus callosum	Present		4D

Structure	Level	Pattern	Image	Note
TS22: cerebral internal capsule	Strong		5A

Structure	Level	Pattern	Image	Note
TS25: cerebral internal capsule	Strong		5B

Structure	Level	Pattern	Image	Note
TS27: cerebral internal capsule	Strong		5C

Structure	Level	Pattern	Image	Note
TS28: cerebral internal capsule	Strong		5D