Probe:	Cux2 cDNA
Probe preparation:	Antisense labelled with digoxigenin RNA

Structure	Level	Pattern	Image	Note
TS17: telencephalon	Weak

Structure	Level	Pattern	Image	Note
TS19: telencephalon	Present	Regionally restricted	1A' , 1A	Expression was detected in the pallium, the subpallium, and the cortical hem. In the subpallium, most expression was localized in the medial ganglionic eminence, with sparse cells in the lateral ganglionic eminence. In the pallium, expression was found in the subventricular zone.

Structure	Level	Pattern	Image	Note
TS20: telencephalon	Present	Regionally restricted	1B' , 1B	Expression was detected in the pallium, the subpallium, and the cortical hem.
TS20: medial ganglionic eminence	Present		1B' , 1B
TS20: lateral ganglionic eminence	Present	Single cells	1B' , 1B
TS20: cerebral cortex subventricular zone	Present		1B , 1B'
TS20: cerebral cortex ventricular layer	Absent		1B' , 1B

Structure	Level	Pattern	Image	Note
TS21: cerebral cortex intermediate zone	Present		1C' , 1C
TS21: cerebral cortex subventricular zone	Present		1C' , 1C
TS21: cerebral cortex marginal layer	Present	Regionally restricted	1C' , 1C	Signal was detected in a row of cells in the marginal zone.
TS21: cortical plate	Present	Single cells	1C' , 1C

Structure	Level	Pattern	Image	Note
TS22: cerebral cortex intermediate zone	Present		1D , 1D'
TS22: cerebral cortex subventricular zone	Present		1D , 1D'
TS22: cerebral cortex marginal layer	Present	Regionally restricted	1D , 1D'	Signal was detected in a row of cells in the marginal zone.
TS22: cortical plate	Present	Single cells	1D' , 1D

Structure	Level	Pattern	Image	Note
TS24: cerebral cortex subventricular zone	Present		1E' , 1E
TS24: cerebral cortex intermediate zone	Present		1E , 1E'
TS24: cerebral cortex marginal layer	Present		1E , 1E'
TS24: cortical plate	Present	Non-Uniform	1E' , 1E	Expression was detected in migrating cells deep in the cortical plate and in condensed cells within the outer-most part of the cortical plate.

Structure	Level	Pattern	Image	Note
TS26: cerebral cortex intermediate zone	Weak		1F' , 1F
TS26: cerebral cortex subventricular zone	Weak		1F' , 1F
TS26: cortical plate	Present	Non-Uniform	1F , 1F'	The majority of the labeled cell population was found in the outer-most part of the cortical plate.

Structure	Level	Pattern	Image	Note
TS28: cortical layer II	Present		1G , 1G'	The majority of labeled cells were detected in the upper part of the cortex (layers II-IV).
TS28: cortical layer III	Present		1G , 1G'	The majority of labeled cells were detected in the upper part of the cortex (layers II-IV).
TS28: cortical layer IV	Present		1G , 1G'	The majority of labeled cells were detected in the upper part of the cortex (layers II-IV).
TS28: cortical layer V	Present	Single cells	1G , 1G'	Some individual labeled cells were identifiable in the deep part of the cortex (layers V-VI).
TS28: cortical layer VI	Present	Single cells	1G , 1G'	Some individual labeled cells were identifiable in the deep part of the cortex (layers V-VI).

Structure	Level	Pattern	Image	Note
TS22: cerebral cortex marginal layer	Present		2A	Staining was comparable to Dlx5 staining.
TS22: cerebral cortex intermediate zone	Present		2A	Staining in the subventricular zone and intermediate zone far exceeded that of the Dlx5 population.
TS22: cerebral cortex subventricular zone	Present		2A	Staining in the subventricular zone and intermediate zone far exceeded that of the Dlx5 population.

Structure	Level	Pattern	Image	Note
TS24: cerebral cortex marginal layer	Present		2B	Staining was comparable to Dlx5 staining.
TS24: cerebral cortex intermediate zone	Present		2B	Staining in the subventricular zone and intermediate zone far exceeded that of the Dlx5 population.
TS24: cerebral cortex subventricular zone	Present		2B	Staining in the subventricular zone and intermediate zone far exceeded that of the Dlx5 population.
TS24: cortical plate	Present	Regionally restricted	2B	Individual cells were found in the innermost part of the cortical plate, whereas staining accumulated in the outermost part of the cortical plate.

Structure	Level	Pattern	Image	Note
TS26: cerebral cortex marginal zone	Present		2C	Staining was comparable to Dlx5 staining.
TS26: cerebral cortex intermediate zone	Present		2C	Staining in the subventricular zone and intermediate zone far exceeded that of the Dlx5 population.
TS26: cerebral cortex subventricular zone	Present		2C	Staining in the subventricular zone and intermediate zone far exceeded that of the Dlx5 population.
TS26: cortical plate	Present	Regionally restricted	2C	The majority of the Cux2-positive population was located below the marginal zone in the position of the future upper cortical layers.

Structure	Level	Pattern	Image	Note
TS23: cerebral cortex marginal layer	Present	Regionally restricted	2G , 2I	Cux2-positive/DLX-positive cells were noted in the marginal zone.
TS23: cerebral cortex intermediate zone	Present	Regionally restricted	2H , 2I	All DLX-positive cells were Cux2-positive, but the majority of cells in the subventricular zone and intermediate zone were Cux2-positive;DLX-negative.
TS23: cerebral cortex subventricular zone	Present	Regionally restricted	2H , 2I	All DLX-positive cells were Cux2-positive, but the majority of cells in the subventricular zone and intermediate zone were Cux2-positive;DLX-negative.
TS23: cerebral cortex ventricular layer	Present	Single cells	2I

Structure	Level	Pattern	Image	Note
TS23: cerebral cortex	Present	Regionally restricted	2J	The number of Cux2-positive;DLX-positive cells was reduced in the mutant cortex.
TS23: cerebral cortex intermediate zone	Present		2J	The number of Cux2-positive;DLX-negative cells was not affected in the mutant.
TS23: cerebral cortex subventricular zone	Present		2J	The number of Cux2-positive;DLX-negative cells was not affected in the mutant.

Structure	Level	Pattern	Image	Note
TS21: cerebral cortex intermediate zone	Present		3A
TS21: cerebral cortex subventricular zone	Present		3A
TS21: cerebral cortex marginal layer	Present	Regionally restricted	3A	Expression was detected in a row of cells in the marginal zone.
TS21: cerebral cortex ventricular layer	Present	Single cells	3A	Individually expressing cells correspond to migrating interneurons.
TS21: cortical plate	Present	Single cells	3A	Individually expressing cells correspond to migrating interneurons.

Structure	Level	Pattern	Image	Note
TS26: cerebral cortex intermediate zone	Present		3D
TS26: cerebral cortex subventricular zone	Present		3D
TS26: cortical plate	Present	Regionally restricted	3D	Expression was detected in cells migrating through the deep part of the cortical plate (which will give rise to cortical layers V and VI) and in condensed cells in the outer-most part of the cortical plate (which will give rise to cortical layers II-IV).

Structure	Level	Pattern	Image	Note
TS28: cortical layer II	Present		3G	Expression was detected in neurons as indicated by the double labeling with Rbfox3.
TS28: cortical layer III	Present		3G	Expression was detected in neurons as indicated by the double labeling with Rbfox3.
TS28: cortical layer IV	Present		3G	Expression was detected in neurons as indicated by the double labeling with Rbfox3.

Structure	Level	Pattern	Image	Note
TS28: cerebral cortex	Present		3J , 3I , 3K	Coexpression was noted with the pan-neuronal marker Rbfox3. Cux2 was expressed in upper layer neurons as well as in individual neurons distributed in the deep layers of the cortex.

Structure	Level	Pattern	Image	Note
TS28: cerebral cortex	Present		3L , 3M , 3N	Cux2+;DLX+ co-labeled cells were identified as interneurons and distributed throughout all the cortical layers.

Structure	Level	Pattern	Image	Note
TS26: cortical plate	Present	Regionally restricted	3O	Expression accumulated in the upper part of the cortical plate.

Structure	Level	Pattern	Image	Note
TS26: cerebral cortex	Present	Regionally restricted	3P , 3Q	In the Reeler cortex, expression remained in deep positions and appeared more diffuse and dispersed compared to wild type. Cux2-positive cells localized in superficial positions in the Reeler cortex coexpressed DLX and thus represent cortical interneurons.

Structure	Level	Pattern	Image	Note
TS20: cerebral cortex subventricular zone	Present		4B , 4A	Many Cux2-positive cells in the subventricular zone were also labeled with Pcna.

Structure	Level	Pattern	Image	Note
TS20: cerebral cortex subventricular zone	Present		4F , 4E	There was co-expression of Cux2 and PHH3, which labels cells in late G2 and M-phase.

Structure	Level	Pattern	Image	Note
TS20: cerebral cortex subventricular zone	Present		4G , 4H	After a 6-hour pulse of BrdU, a subfraction of Cux2-positive cells showed BrdU labelling.

Structure	Level	Pattern	Image	Note
TS21: cerebral cortex mantle layer	Present		5B
TS21: cerebral cortex subventricular zone	Present		5B

Structure	Level	Pattern	Image	Note
TS21: cerebral cortex mantle layer	Present		5C
TS21: cerebral cortex subventricular zone	Present		5C

Structure	Level	Pattern	Image	Note
TS21: cerebral cortex subventricular zone	Present		5E	There was a considerable decrease in Cux2-positive cells in the mutant compared with wild type.
TS21: cerebral cortex marginal layer	Present		5E	Cux2+/DLX+ cells were still identifiable in the marginal zone.

Structure	Level	Pattern	Image	Note
TS21: cerebral cortex subventricular zone	Present		5F	There was a considerable decrease in Cux2-positive cells in the mutant compared with wild type.
TS21: cerebral cortex mantle layer	Present		5F

Structure	Level	Pattern	Image	Note
TS23: cerebral cortex intermediate zone	Present		5H
TS23: cerebral cortex marginal layer	Present		5H
TS23: cerebral cortex subventricular zone	Present		5H

Structure	Level	Pattern	Image	Note
TS23: cerebral cortex intermediate zone	Present		5K , 5J	There was a considerable decrease in Cux2-positive cells in the mutant compared with wild type. The Cux2-positive population of upper layer neurons was lost from the subventricular zone and underrepresented in the intermediate zone.
TS23: cerebral cortex subventricular zone	Present		5K , 5J	There was a considerable decrease in Cux2-positive cells in the mutant compared with wild type. The Cux2-positive population of upper layer neurons was lost from the subventricular zone and underrepresented in the intermediate zone.
TS23: cerebral cortex marginal layer	Present		5K , 5J	The Cux2+/DLX+ interneuron population in the marginal zone was increased in the mutant.

Structure	Level	Pattern	Image	Note
TS25: cerebral cortex intermediate zone	Present		5M
TS25: cerebral cortex marginal zone	Present		5M
TS25: cerebral cortex subventricular zone	Present		5M
TS25: cortical plate	Present		5M

Structure	Level	Pattern	Image	Note
TS25: cerebral cortex intermediate zone	Present		5N
TS25: cerebral cortex marginal zone	Present		5N
TS25: cerebral cortex subventricular zone	Present		5N
TS25: cortical plate	Present		5N

Structure	Level	Pattern	Image	Note
TS25: cerebral cortex marginal zone	Present		5P	The mutant cortex showed an increase in Cux2+/DLX+ interneurons in the marginal zone.
TS25: cerebral cortex intermediate zone	Trace	Single cells	5P	Cux2-positive cells are almost absent.
TS25: cerebral cortex subventricular zone	Trace	Single cells	5P	Cux2-positive cells are almost absent.
TS25: cortical plate	Present		5P	Cux2-positive cells are less numerous in the mutant cortical plate compared to wild type.

Structure	Level	Pattern	Image	Note
TS25: cerebral cortex marginal zone	Present		5Q	The mutant cortex showed an increase in Cux2+/DLX+ interneurons in the marginal zone.
TS25: cerebral cortex intermediate zone	Trace	Single cells	5Q	Cux2-positive cells are almost absent.
TS25: cerebral cortex subventricular zone	Trace	Single cells	5Q	Cux2-positive cells are almost absent.
TS25: cortical plate	Present		5Q	Cux2-positive cells are less numerous in the mutant cortical plate compared to wild type.