Probe:	IMAGE clone 4979208
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	Very strong	Non-Uniform	Spry2-Shh E11.5_coronal	There is very strong staining in the MZ of the dorsal thalamus & prethalamus. Staining is either absent or ubiquitous in the MZ of the dorsal & ventral POAs, the PVH, VMH & ARH nuclei, the AVH, anterior & posterior IDs, premam, mam & supramam zones & TT.
TS19: diencephalon lateral wall ventricular layer	Very strong	Non-Uniform	Spry2-Shh E11.5_coronal	There is very strong staining in the VZ of the dorsal thalamus, prethalamus & ventral POA. Staining is either absent or ubiquitous in the VZ of the dorsal POA, PVH, VMH & ARH nuclei, the AVH, anterior & posterior IDs, premam, mam & supramam zones & TT.
TS19: telencephalon mantle layer	Ambiguous		Spry2-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		Spry2-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		Spry2-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	Ambiguous		Spry2-Shh E12.5_coronal	Staining is either absent or ubiquitous in the ventricular zone of the cerebral cortex.
TS20: cortical plate	Ambiguous		Spry2-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	Spry2-Shh E12.5_coronal	There is moderate 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	Ambiguous		Spry2-Shh E12.5_coronal	Staining is either absent or ubiquitous in the mantle zone of the dorsal and ventral POAs, PVH nucleus and the AVH.
TS20: hypothalamus ventricular layer	Very strong	Non-Uniform	Spry2-Shh E12.5_coronal	There is very strong staining in the ventricular zone of the ventral POA and the AVH and weak, scattered staining in the PVH nucleus. Staining is either absent or ubiquitous in the ventricular zone of the dorsal POA.
TS20: thalamus mantle layer	Ambiguous		Spry2-Shh E12.5_coronal	Staining is either absent or ubiquitous in the mantle zone of the dorsal thalamus and prethalamus.
TS20: thalamus ventricular layer	Moderate		Spry2-Shh E12.5_coronal	There is moderate staining in the ventricular zone of the dorsal thalamus and prethalamus.

Structure	Level	Pattern	Image	Note
TS22: cerebral cortex intermediate zone	Ambiguous		Spry2-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	Ambiguous		Spry2-Shh E14.5_coronal	Staining is either absent or ubiquitous in the ventricular zone of the cerebral cortex.
TS22: cortical plate	Ambiguous		Spry2-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	Ambiguous		Spry2-Shh E14.5_coronal	Staining is either absent or ubiquitous in the mantle zone of the premam, mam and supramam zones and the TT.
TS22: diencephalon ventricular layer	Moderate	Non-Uniform	Spry2-Shh E14.5_coronal	There is moderate staining in the ventricular zone of the TT. Staining is either absent or ubiquitous in the ventricular zone of the premam, mam and supramam zones.
TS22: ganglionic eminence	Moderate	Non-Uniform	Spry2-Shh E14.5_coronal	There is moderate 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	Ambiguous		Spry2-Shh E14.5_coronal	Staining is either absent or ubiquitous in the mantle zone of the dorsal and ventral POAs, the PVH, VMH and ARH nuclei, the AVH and anterior and posterior IDs.
TS22: hypothalamus ventricular layer	Moderate	Non-Uniform	Spry2-Shh E14.5_coronal	There is moderate staining in the VZ of the anterior and posterior IDs and weak, scattered staining in the dorsal and ventral POAs, PVH nucleus and the AVH. Staining is either absent or ubiquitous in the VZ of the VMH and ARH nuclei.
TS22: thalamus mantle layer	Moderate	Non-Uniform	Spry2-Shh E14.5_coronal	There is moderate 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	Weak		Spry2-Shh E14.5_coronal	There is weak staining in the ventricular zone of the dorsal thalamus and prethalamus.

Structure	Level	Pattern	Image	Note
TS24: cerebral cortex intermediate zone	Weak		Spry2-Shh E16.5_coronal	There is weak staining in the postmitotic neurons in the intermediate zone of the cerebral cortex.
TS24: cerebral cortex ventricular layer	Weak		Spry2-Shh E16.5_coronal	There is weak staining in the ventricular zone of the cerebral cortex.
TS24: cortical plate	Weak		Spry2-Shh E16.5_coronal	There is weak staining in the postmitotic neurons in the cortical plate of the cerebral cortex.
TS24: diencephalon mantle layer	Moderate	Non-Uniform	Spry2-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		Spry2-Shh E16.5_coronal	Staining is either absent or ubiquitous in the ventricular zone of the premam, mam and supramam zones.
TS24: ganglionic eminence	Ambiguous		Spry2-Shh E16.5_coronal	Staining is either absent or ubiquitous in the ventricular and mantle zones of the ganglionic eminences.
TS24: hypothalamus mantle layer	Ambiguous		Spry2-Shh E16.5_coronal	Staining is either absent or ubiquitous in the mantle zone of the dorsal and ventral POAs, the PVH, SCH, DMH, VMH and ARH nuclei, the AVH, the DAH and the PH.
TS24: hypothalamus ventricular layer	Strong	Non-Uniform	Spry2-Shh E16.5_coronal	There is strong staining in the ventricular zone of the PVH, SCH and DMH nuclei, the AVH and the DAH and weak staining in the dorsal and ventral POAs. Staining is either absent or ubiquitous in the ventricular zone of the VMH and ARH nuclei and the PH.
TS24: thalamus mantle layer	Moderate	Non-Uniform	Spry2-Shh E16.5_coronal	There is moderate staining in the mantle zone of the dorsal thalamus. Staining is either absent or ubiquitous in the mantle zone of the prethalamus.
TS24: thalamus ventricular layer	Strong		Spry2-Shh E16.5_coronal	There is strong staining in the ventricular zone of the dorsal thalamus and prethalamus.