Probe:	UI-M-BH2.3-aoj-b-09-0-UI.s1
Probe preparation:	Antisense labelled with digoxigenin RNA
Visualized with:	Alkaline phosphatase
Assay notes:	These specimens were also hybridized with a fluorescein-labeled Shh probe MGI:2110655; detected with TBT/BCIP. The results are reported in Supp. Table 4, whose legend includes a key to the abbreviations used and more fully details the regions examined.

Structure	Level	Pattern	Image	Note
TS19: diencephalon lateral wall mantle layer	Moderate	Non-Uniform	Sema5a-Shh E11.5_coronal	Staining is moderate in the MZ of the dorsal thalamus & weak in the AVH. Staining is either absent or ubiquitous in the MZ of the prethalamus, dorsal & ventral POAs, PVH, VMH & ARH nuclei, anterior & posterior IDs, premam, mam & supramam zones & TT.
TS19: diencephalon lateral wall ventricular layer	Ambiguous		Sema5a-Shh E11.5_coronal	Staining is either absent or ubiquitous in the ventricular zone of the dorsal thalamus, the prethalamus, dorsal and ventral POAs, the PVH, VMH and ARH nuclei, the AVH, the anterior and posterior IDs, the premam, mam and supramam zones and the TT.
TS19: telencephalon mantle layer	Ambiguous		Sema5a-Shh E11.5_coronal	Staining is either absent or ubiquitous in the postmitotic neurons in the intermediate zone and cortical plate of the cerebral cortex and in the mantle zone of the ganglionic eminences.
TS19: telencephalon ventricular layer	Very strong	Non-Uniform	Sema5a-Shh E11.5_coronal	There is very strong staining in the ventricular zone of the cerebral cortex and moderate staining in the ganglionic eminences.

Structure	Level	Pattern	Image	Note
TS20: cerebral cortex intermediate zone	Ambiguous		Sema5a-Shh E12.5_coronal	Staining is either absent or ubiquitous in the postmitotic neurons in the intermediate zone of the cerebral cortex.
TS20: cerebral cortex ventricular layer	Very strong		Sema5a-Shh E12.5_coronal	There is very strong staining in the ventricular zone of the cerebral cortex.
TS20: cortical plate	Ambiguous		Sema5a-Shh E12.5_coronal	Staining is either absent or ubiquitous in the postmitotic neurons in the cortical plate of the cerebral cortex.
TS20: ganglionic eminence	Moderate	Non-Uniform	Sema5a-Shh E12.5_coronal	There is moderate, scattered staining in the ventricular zone of the ganglionic eminences. Staining is either absent or ubiquitous in the mantle zone of the ganglionic eminences.
TS20: hypothalamus mantle layer	Weak	Non-Uniform	Sema5a-Shh E12.5_coronal	There is weak staining in the mantle zone of the PVH nucleus. Staining is either absent or ubiquitous in the mantle zone of the dorsal and ventral POAs and the AVH.
TS20: hypothalamus ventricular layer	Weak	Non-Uniform	Sema5a-Shh E12.5_coronal	There is weak staining in the ventricular zone of the AVH. Staining is either absent or ubiquitous in the ventricular zone of the dorsal and ventral POAs and the PVH nucleus.
TS20: thalamus mantle layer	Very strong	Non-Uniform	Sema5a-Shh E12.5_coronal	There is very strong staining in the mantle zone of the dorsal thalamus. Staining is either absent or ubiquitous in the mantle zone of the prethalamus.
TS20: thalamus ventricular layer	Weak	Non-Uniform	Sema5a-Shh E12.5_coronal	There is weak staining in the ventricular zone of the prethalamus. Staining is either absent or ubiquitous in the ventricular zone of the dorsal thalamus.

Structure	Level	Pattern	Image	Note
TS22: cerebral cortex intermediate zone	Ambiguous		Sema5a-Shh E14.5_coronal	Staining is either absent or ubiquitous in the postmitotic neurons in the intermediate zone of the cerebral cortex.
TS22: cerebral cortex ventricular layer	Very strong		Sema5a-Shh E14.5_coronal	There is very strong staining in the ventricular zone of the cerebral cortex.
TS22: cortical plate	Ambiguous		Sema5a-Shh E14.5_coronal	Staining is either absent or ubiquitous in the postmitotic neurons in the cortical plate of the cerebral cortex.
TS22: diencephalon mantle layer	Weak	Non-Uniform	Sema5a-Shh E14.5_coronal	There is weak staining in the mantle zone of the premam zone. Staining is either absent or ubiquitous in the mantle zone of the TT and mam and supramam zones.
TS22: diencephalon ventricular layer	Ambiguous		Sema5a-Shh E14.5_coronal	Staining is either absent or ubiquitous in the ventricular zone of the premam, mam and supramam zones and the TT.
TS22: ganglionic eminence	Weak	Non-Uniform	Sema5a-Shh E14.5_coronal	There is weak staining in the ventricular zone of the ganglionic eminences. Staining is either absent or ubiquitous in the mantle zone of the ganglionic eminences.
TS22: hypothalamus mantle layer	Weak	Non-Uniform	Sema5a-Shh E14.5_coronal	There is weak staining in the mantle zone of the PVH nucleus. Staining is either absent or ubiquitous in the mantle zone of the dorsal and ventral POAs, the VMH and ARH nuclei, the AVH and anterior and posterior IDs.
TS22: hypothalamus ventricular layer	Very strong	Non-Uniform	Sema5a-Shh E14.5_coronal	There is very strong staining in the ventricular zone of the AVH. Staining is either absent or ubiquitous in the ventricular zone of the dorsal and ventral POAs, the PVH, VMH and ARH nuclei and anterior and posterior IDs.
TS22: thalamus mantle layer	Very strong	Non-Uniform	Sema5a-Shh E14.5_coronal	There is very strong staining in the mantle zone of the dorsal thalamus. Staining is either absent or ubiquitous in the mantle zone of the prethalamus.
TS22: thalamus ventricular layer	Moderate	Non-Uniform	Sema5a-Shh E14.5_coronal	There is moderate staining in the ventricular zone of the prethalamus. Staining is either absent or ubiquitous in the ventricular zone of the dorsal thalamus.

Structure	Level	Pattern	Image	Note
TS24: cerebral cortex intermediate zone	Ambiguous		Sema5a-Shh E16.5_coronal	Staining is either absent or ubiquitous in the postmitotic neurons in the intermediate zone of the cerebral cortex.
TS24: cerebral cortex ventricular layer	Very strong		Sema5a-Shh E16.5_coronal	There is very strong staining in the ventricular zone of the cerebral cortex.
TS24: cortical plate	Ambiguous		Sema5a-Shh E16.5_coronal	Staining is either absent or ubiquitous in the postmitotic neurons in the cortical plate of the cerebral cortex.
TS24: diencephalon mantle layer	Moderate	Non-Uniform	Sema5a-Shh E16.5_coronal	There is moderate staining in the mantle zone of the premam zone. Staining is either absent or ubiquitous in the mantle zone of the mam and supramam zones.
TS24: diencephalon ventricular layer	Ambiguous		Sema5a-Shh E16.5_coronal	Staining is either absent or ubiquitous in the ventricular zone of the premam, mam and supramam zones.
TS24: ganglionic eminence	Moderate	Non-Uniform	Sema5a-Shh E16.5_coronal	There is moderate staining in the mantle zone and weak staining in the ventricular zone of the ganglionic eminences.
TS24: hypothalamus mantle layer	Very strong	Non-Uniform	Sema5a-Shh E16.5_coronal	There is very strong staining in the MZ of the PVH nucleus and the AVH and weak staining in the dorsal and ventral POAs and the VMH nucleus. Staining is either absent or ubiquitous in the MZ of the DAH, the SCH, DMH and ARH nuclei and the PH.
TS24: hypothalamus ventricular layer	Ambiguous		Sema5a-Shh E16.5_coronal	Staining is either absent or ubiquitous in the ventricular zone of the dorsal and ventral POAs, the PVH, SCH, DMH, VMH and ARH nuclei, the AVH, the DAH and the PH.
TS24: thalamus mantle layer	Strong		Sema5a-Shh E16.5_coronal	There is strong staining in the mantle zone of the dorsal thalamus and prethalamus.
TS24: thalamus ventricular layer	Ambiguous		Sema5a-Shh E16.5_coronal	Staining is either absent or ubiquitous in the ventricular zone of the dorsal thalamus and prethalamus.