Probe:	UI-M-BH1-amw-c-06-0-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	Strong	Non-Uniform	Gnao1-Shh E11.5-coronal	There is strong staining in the MZ of the prethalamus. Staining is either absent or ubiquitous in the MZ of the dorsal thalamus, dorsal & ventral POAs, the PVH, VMH & ARH nuclei, the AVH, anterior & posterior IDs, premam, mam & supramam zones & the TT.
TS19: diencephalon lateral wall ventricular layer	Strong	Non-Uniform	Gnao1-Shh E11.5-coronal	There is strong staining in the VZ of the dorsal POA, PVH nucleus, anterior & posterior IDs, TT & mam zone. Staining is either absent or ubiquitous in the VZ of dorsal thalamus, prethalamus, ventral POA, AVH, VMH & ARH nuclei & premam & supramam zones.
TS19: telencephalon mantle layer	Ambiguous		Gnao1-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	Ambiguous		Gnao1-Shh E11.5-coronal	Staining is either absent or ubiquitous in the ventricular zone of the cerebral cortex and the ganglionic eminences.

Structure	Level	Pattern	Image	Note
TS20: cerebral cortex intermediate zone	Ambiguous		Gnao1-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	Weak		Gnao1-Shh E12.5-coronal	There is weak staining in the ventricular zone of the cerebral cortex.
TS20: cortical plate	Ambiguous		Gnao1-Shh E12.5-coronal	Staining is either absent or ubiquitous in the postmitotic neurons in the cortical plate of the cerebral cortex.
TS20: diencephalon mantle layer	Moderate		Gnao1-Shh E12.5-coronal	There is moderate staining in the mantle zone of the premam, mam and supramam zones and the TT.
TS20: diencephalon ventricular layer	Ambiguous		Gnao1-Shh E12.5-coronal	Staining is either absent or ubiquitous in the ventricular zone of the premam, mam and supramam zones and the TT.
TS20: ganglionic eminence	Strong	Non-Uniform	Gnao1-Shh E12.5-coronal	There is strong staining in the ventricular zone of the ganglionic eminences and weak staining in the mantle zone of the ganglionic eminences.
TS20: hypothalamus mantle layer	Very strong	Non-Uniform	Gnao1-Shh E12.5-coronal	There is very strong staining in the mantle zone of the anterior and posterior IDs, strong staining in the dorsal and ventral POAs, the PVH nucleus and the AVH and moderate staining in the VMH and ARH nuclei.
TS20: hypothalamus ventricular layer	Ambiguous		Gnao1-Shh E12.5-coronal	Staining is either absent or ubiquitous in the ventricular zone of the dorsal and ventral POAs, the PVH, VMH and ARH nuclei, the AVH and anterior and posterior IDs.
TS20: thalamus mantle layer	Moderate		Gnao1-Shh E12.5-coronal	There is moderate staining in the mantle zone of the dorsal thalamus and prethalamus.
TS20: thalamus ventricular layer	Ambiguous		Gnao1-Shh E12.5-coronal	Staining is either absent or ubiquitous in the ventricular zone of the dorsal thalamus and prethalamus.

Structure	Level	Pattern	Image	Note
TS22: cerebral cortex intermediate zone	Moderate		Gnao1-Shh E14.5-coronal	There is moderate staining in the postmitotic neurons in the intermediate zone of the cerebral cortex.
TS22: cerebral cortex ventricular layer	Very strong		Gnao1-Shh E14.5-coronal	There is very strong staining in the ventricular zone of the cerebral cortex.
TS22: cortical plate	Moderate		Gnao1-Shh E14.5-coronal	There is moderate staining in the postmitotic neurons in the cortical plate of the cerebral cortex.
TS22: diencephalon mantle layer	Moderate	Non-Uniform	Gnao1-Shh E14.5-coronal	There is moderate staining in the mantle zone of the premam zone and the TT. Staining is either absent or ubiquitous in the mantle zone of the mam and supramam zones.
TS22: diencephalon ventricular layer	Very strong		Gnao1-Shh E14.5-coronal	There is very strong staining in the ventricular zone of the premam, mam and supramam zones and the TT.
TS22: ganglionic eminence	Very strong	Non-Uniform	Gnao1-Shh E14.5-coronal	There is very strong staining in the ventricular zone of the ganglionic eminences and weak staining in the mantle zone of the ganglionic eminences.
TS22: hypothalamus mantle layer	Weak	Non-Uniform	Gnao1-Shh E14.5-coronal	There is weak staining in the mantle zone of the dorsal and ventral POAs and the AVH. Staining is either absent or ubiquitous in the mantle zone of the VMH and ARH nuclei, the AVH and anterior and posterior IDs.
TS22: hypothalamus ventricular layer	Very strong	Non-Uniform	Gnao1-Shh E14.5-coronal	There is very strong staining in the VZ of the dorsal POA, strong staining in the VMH and ARH nuclei, the AVH and anterior and posterior IDs and weak staining in the PVH nucleus. Staining is either absent or ubiquitous in the VZ of the ventral POA.
TS22: thalamus mantle layer	Moderate	Non-Uniform	Gnao1-Shh E14.5-coronal	There is moderate staining in the mantle zone of the dorsal thalamus and weak staining in the prethalamus.
TS22: thalamus ventricular layer	Very strong		Gnao1-Shh E14.5-coronal	There is very strong staining in the ventricular zone of the dorsal thalamus and prethalamus.

Structure	Level	Pattern	Image	Note
TS24: cerebral cortex intermediate zone	Ambiguous		Gnao1-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		Gnao1-Shh E16.5-coronal	There is very strong staining in the ventricular zone of the cerebral cortex.
TS24: cortical plate	Ambiguous		Gnao1-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	Weak	Non-Uniform	Gnao1-Shh E16.5-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 mam and supramam zones.
TS24: diencephalon ventricular layer	Very strong		Gnao1-Shh E16.5-coronal	There is very strong staining in the ventricular zone of the premam, mam and supramam zones.
TS24: ganglionic eminence	Very strong	Non-Uniform	Gnao1-Shh E16.5-coronal	There is very strong staining in the ventricular zone of the ganglionic eminences. Staining is either absent or ubiquitous in the mantle zone of the ganglionic eminences.
TS24: hypothalamus mantle layer	Weak	Non-Uniform	Gnao1-Shh E16.5-coronal	There is weak staining in the mantle zone of the SCH nucleus and the PH. Staining is either absent or ubiquitous in the mantle zone of the dorsal and ventral POAs, the PVH, DMH, VMH and ARH nuclei, the AVH and the DAH.
TS24: hypothalamus ventricular layer	Very strong		Gnao1-Shh E16.5-coronal	There is very strong staining 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	Weak		Gnao1-Shh E16.5-coronal	There is weak staining in the mantle zone of the dorsal thalamus and prethalamus.
TS24: thalamus ventricular layer	Very strong		Gnao1-Shh E16.5-coronal	There is very strong staining in the ventricular zone of the dorsal thalamus and prethalamus.