Probe:	MGC clone 90675
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		Insm1-Shh E11.5-sagittal , Insm1-Shh E11.5-coronal	There is moderate staining in the mantle zone of the dorsal thalamus, 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: diencephalon lateral wall ventricular layer	Ambiguous		Insm1-Shh E11.5-sagittal , Insm1-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	Moderate	Non-Uniform	Insm1-Shh E11.5-coronal , Insm1-Shh E11.5-sagittal	There is moderate staining in the mantle zone of the ganglionic eminences. Staining is either absent or ubiquitous in the postmitotic neurons in the intermediate zone and cortical plate of the cerebral cortex.
TS19: telencephalon ventricular layer	Ambiguous		Insm1-Shh E11.5-coronal , Insm1-Shh E11.5-sagittal	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	Strong		Insm1-Shh E12.5-coronal , Insm1-Shh E12.5-sagittal	There is strong staining in the postmitotic neurons in the intermediate zone of the cerebral cortex.
TS20: cerebral cortex ventricular layer	Ambiguous		Insm1-Shh E12.5-coronal , Insm1-Shh E12.5-sagittal	Staining is either absent or ubiquitous in the ventricular zone of the cerebral cortex.
TS20: cortical plate	Strong		Insm1-Shh E12.5-coronal , Insm1-Shh E12.5-sagittal	There is strong staining in the postmitotic neurons in the cortical plate of the cerebral cortex.
TS20: diencephalon mantle layer	Strong		Insm1-Shh E12.5-sagittal , Insm1-Shh E12.5-coronal	There is strong staining in the mantle zone of the premam, mam and supramam zones and the TT.
TS20: diencephalon ventricular layer	Ambiguous		Insm1-Shh E12.5-coronal , Insm1-Shh E12.5-sagittal	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	Insm1-Shh E12.5-sagittal , Insm1-Shh E12.5-coronal	There is strong staining in the mantle zone of the ganglionic eminences. Staining is either absent or ubiquitous in the ventricular zone of the ganglionic eminences.
TS20: hypothalamus mantle layer	Strong		Insm1-Shh E12.5-sagittal , Insm1-Shh E12.5-coronal	There is strong staining in the mantle zone of the dorsal and ventral POAs, the PVH, VMH and ARH nuclei, the AVH and anterior and posterior IDs.
TS20: hypothalamus ventricular layer	Ambiguous		Insm1-Shh E12.5-coronal , Insm1-Shh E12.5-sagittal	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	Strong		Insm1-Shh E12.5-coronal , Insm1-Shh E12.5-sagittal	There is strong staining in the mantle zone of the dorsal thalamus and prethalamus.
TS20: thalamus ventricular layer	Ambiguous		Insm1-Shh E12.5-sagittal , Insm1-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		Insm1-Shh E14.5-coronal , Insm1-Shh E14.5-sagittal	There is moderate staining in the postmitotic neurons in the intermediate zone of the cerebral cortex.
TS22: cerebral cortex ventricular layer	Ambiguous		Insm1-Shh E14.5-coronal , Insm1-Shh E14.5-sagittal	Staining is either absent or ubiquitous in the ventricular zone of the cerebral cortex.
TS22: cortical plate	Moderate		Insm1-Shh E14.5-sagittal , Insm1-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		Insm1-Shh E14.5-sagittal , Insm1-Shh E14.5-coronal	There is moderate staining in the mantle zone of the premam, mam and supramam zones and the TT.
TS22: diencephalon ventricular layer	Ambiguous		Insm1-Shh E14.5-coronal , Insm1-Shh E14.5-sagittal	Staining is either absent or ubiquitous in the ventricular zone of the premam, mam and supramam zones and the TT.
TS22: ganglionic eminence	Moderate	Non-Uniform	Insm1-Shh E14.5-sagittal , Insm1-Shh E14.5-coronal	There is moderate staining in the mantle zone of the ganglionic eminences. Staining is either absent or ubiquitous in the ventricular zone of the ganglionic eminences.
TS22: hypothalamus mantle layer	Very strong	Non-Uniform	Insm1-Shh E14.5-sagittal , Insm1-Shh E14.5-coronal	There is very strong staining in the mantle zone of the PVH nucleus and moderate staining in the dorsal and ventral POAs, the VMH and ARH nuclei, the AVH and anterior and posterior IDs.
TS22: hypothalamus ventricular layer	Ambiguous		Insm1-Shh E14.5-coronal , Insm1-Shh E14.5-sagittal	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.
TS22: thalamus mantle layer	Moderate		Insm1-Shh E14.5-coronal , Insm1-Shh E14.5-sagittal	There is moderate staining in the mantle zone of the dorsal thalamus and prethalamus.
TS22: thalamus ventricular layer	Ambiguous		Insm1-Shh E14.5-coronal , Insm1-Shh E14.5-sagittal	Staining is either absent or ubiquitous in the ventricular zone of the dorsal thalamus and prethalamus.

Structure	Level	Pattern	Image	Note
TS24: cerebral cortex intermediate zone	Moderate		Insm1-Shh E16.5-sagittal , Insm1-Shh E16.5-coronal	There is moderate staining in the postmitotic neurons in the intermediate zone of the cerebral cortex.
TS24: cerebral cortex ventricular layer	Ambiguous		Insm1-Shh E16.5-coronal , Insm1-Shh E16.5-sagittal	Staining is either absent or ubiquitous in the ventricular zone of the cerebral cortex.
TS24: cortical plate	Moderate		Insm1-Shh E16.5-coronal , Insm1-Shh E16.5-sagittal	There is moderate staining in the postmitotic neurons in the cortical plate of the cerebral cortex.
TS24: diencephalon mantle layer	Moderate		Insm1-Shh E16.5-coronal , Insm1-Shh E16.5-sagittal	There is moderate staining in the mantle zone of the premam, mam and supramam zones.
TS24: diencephalon ventricular layer	Ambiguous		Insm1-Shh E16.5-sagittal , Insm1-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	Insm1-Shh E16.5-sagittal , Insm1-Shh E16.5-coronal	There is moderate staining in the mantle zone of the ganglionic eminences. Staining is either absent or ubiquitous in the ventricular zone of the ganglionic eminences.
TS24: hypothalamus mantle layer	Very strong	Non-Uniform	Insm1-Shh E16.5-coronal , Insm1-Shh E16.5-sagittal	There is very strong staining in the mantle zone of the PVH and SCH nuclei and moderate staining in the dorsal and ventral POAs, the DMH, VMH and ARH nuclei, the AVH, the DAH and the PH.
TS24: hypothalamus ventricular layer	Ambiguous		Insm1-Shh E16.5-coronal , Insm1-Shh E16.5-sagittal	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	Moderate		Insm1-Shh E16.5-coronal , Insm1-Shh E16.5-sagittal	There is moderate staining in the mantle zone of the dorsal thalamus and prethalamus.
TS24: thalamus ventricular layer	Ambiguous		Insm1-Shh E16.5-sagittal , Insm1-Shh E16.5-coronal	Staining is either absent or ubiquitous in the ventricular zone of the dorsal thalamus and prethalamus.