Probe:	Otx2_riboprobe
Probe preparation:	Antisense labelled with digoxigenin RNA
Visualized with:	Alkaline phosphatase
Assay notes:	The authors analyzed data from the Allen Developing Mouse Brain.

Structure	Level	Pattern	Image	Note
TS19: midbrain	Present		2a
TS19: midbrain mantle layer	Absent		2a

Structure	Level	Pattern	Image	Note
TS19: midbrain	Present		2b
TS19: midbrain mantle layer	Absent		2b

Structure	Level	Pattern	Image	Note
TS19: midbrain	Present		2c
TS19: midbrain mantle layer	Absent		2c

Structure	Level	Pattern	Image	Note
TS19: midbrain	Present		2d
TS19: midbrain mantle layer	Absent		2d

Structure	Level	Pattern	Image	Note
TS19: midbrain	Present		2e
TS19: midbrain mantle layer	Absent		2e

Structure	Level	Pattern	Image	Note
TS19: midbrain	Present		2f
TS19: midbrain mantle layer	Absent		2f

Structure	Level	Pattern	Image	Note
TS22: superior colliculus	Present	Regionally restricted	4a	Sagittal sections showed a positive and elongated dorsorostral area that corresponds to the superior colliculus, which included the still more rostral 'tectal gray' formation.
TS22: inferior colliculus	Absent		4a	Staining limits ventrocaudally with a mantle domain devoid of signal, which represents the inferior colliculus and its rostral continuation in the interstitial bed nucleus of the brachium of the inferior colliculus.
TS22: midbrain	Present	Regionally restricted	4a	Ventrocaudal expression was observed in the preisthmus or mesomere 2 derivative. This domain seemed wider dorsally ('tor') close to the roof plate. The domain thins out ventralwards and displays a sharp boundary with the Otx2-negative isthmus.
TS22: midbrain-hindbrain junction	Absent		4a

Structure	Level	Pattern	Image	Note
TS22: superior colliculus	Present	Regionally restricted	4b	Sagittal sections showed a positive and elongated dorsorostral area that corresponds to the superior colliculus, which included the still more rostral 'tectal gray' formation.
TS22: inferior colliculus	Absent		4b	Staining limits ventrocaudally with a mantle domain devoid of signal, which represents the inferior colliculus and its rostral continuation in the interstitial bed nucleus of the brachium of the inferior colliculus.
TS22: midbrain	Present	Regionally restricted	4b	Ventrocaudal expression was observed in the preisthmus or mesomere 2 derivative. This domain seemed wider dorsally ('tor') close to the roof plate. The domain thins out ventralwards and displays a sharp boundary with the Otx2-negative isthmus.
TS22: midbrain-hindbrain junction	Absent		4b

Structure	Level	Pattern	Image	Note
TS22: superior colliculus	Present		4c
TS22: inferior colliculus	Absent		4c
TS22: midbrain	Present	Regionally restricted	4c	Expression was detected in the preisthmus.

Structure	Level	Pattern	Image	Note
TS22: superior colliculus	Present		4d
TS22: inferior colliculus	Absent		4d
TS22: midbrain	Present	Regionally restricted	4d	Expression was detected in the preisthmus.
TS22: midbrain-hindbrain junction	Absent		4d

Structure	Level	Pattern	Image	Note
TS22: superior colliculus	Present		4e
TS22: inferior colliculus	Absent		4e
TS22: midbrain	Present	Regionally restricted	4e	Expression was detected in the preisthmus.
TS22: midbrain-hindbrain junction	Absent		4e

Structure	Level	Pattern	Image	Note
TS22: superior colliculus	Present		4f
TS22: inferior colliculus	Absent		4f
TS22: midbrain	Present	Regionally restricted	4f	Expression was detected in the preisthmus.
TS22: midbrain-hindbrain junction	Absent		4f

Structure	Level	Pattern	Image	Note
TS22: superior colliculus	Present		4g
TS22: inferior colliculus	Absent		4g
TS22: midbrain	Present	Regionally restricted	4g	Expression was detected in the preisthmus.
TS22: midbrain-hindbrain junction	Absent		4g

Structure	Level	Pattern	Image	Note
TS23: superior colliculus	Present		7i
TS23: midbrain	Present	Regionally restricted	7i	Expression was detected in the preisthmus.

Structure	Level	Pattern	Image	Note
TS23: superior colliculus	Present		7j
TS23: midbrain	Present	Regionally restricted	7j	Expression was detected in the preisthmus.

Structure	Level	Pattern	Image	Note
TS23: superior colliculus	Present		7k
TS23: midbrain	Present	Regionally restricted	7k	Expression was detected in the preisthmus.
TS23: midbrain ventricular layer	Present	Regionally restricted	7k	Expression was detected in the ventricular zone of the inferior colliculus.

Structure	Level	Pattern	Image	Note
TS23: superior colliculus	Present		7l
TS23: midbrain	Present	Regionally restricted	7l	A paramedian parasagittal section illustrated the Otx2 signal in the basal preisthmus, representing a quite narrow transverse and radial interstice placed in a retrorubral position.
TS23: midbrain ventricular layer	Present	Regionally restricted	7l	Expression was detected in the ventricular zone of the inferior colliculus.

Structure	Level	Pattern	Image	Note
TS23: midbrain	Present	Regionally restricted	7m	Dorsoventral horizontal section illustrated the wide dorsal extent of the positive alar preisthmic domain.
TS23: inferior colliculus	Absent		7m

Structure	Level	Pattern	Image	Note
TS23: midbrain	Present	Regionally restricted	7n	More ventrally, the alar preisthmus gets reduced to a narrower Otx2-positive band that underlines the caudal aspect of the inferior colliculus and limits caudally with the negative isthmus domain.
TS23: midbrain-hindbrain junction	Absent		7n

Structure	Level	Pattern	Image	Note
TS23: midbrain	Present	Regionally restricted	7o	More ventrally, the alar preisthmus gets reduced to a narrower Otx2-positive band that underlines the caudal aspect of the inferior colliculus and limits caudally with the negative isthmus domain.
TS23: midbrain-hindbrain junction	Absent		7o

Structure	Level	Pattern	Image	Note
TS23: midbrain	Present	Regionally restricted	7p	More ventrally, the alar preisthmus gets reduced to a narrower Otx2-positive band that underlines the caudal aspect of the inferior colliculus and limits caudally with the negative isthmus domain.
TS23: midbrain-hindbrain junction	Absent		7p

Structure	Level	Pattern	Image	Note
TS26: superior colliculus	Present	Regionally restricted	9a	Labeling was restricted to intermediate and superficial cell layers, plus the ventricular zone.
TS26: midbrain	Present	Regionally restricted	9a	Expression was detected in the preisthmus.
TS26: inferior colliculus	Present	Regionally restricted	9a	The largely negative inferior colliculus/brachium of the inferior colliculus complex only appears labeled at its ventricular zone.
TS26: periaqueductal grey	Absent		9a

Structure	Level	Pattern	Image	Note
TS26: superior colliculus	Present	Regionally restricted	9b	Labeling was restricted to intermediate and superficial cell layers, plus the ventricular zone.
TS26: midbrain	Present	Regionally restricted	9b	Expression was detected in the preisthmus. The dorsalmost Otx2 section illustrates dorsomedial continuity of the positive preisthmic mantle with symmetric bulges at the dorsocaudal midbrain midline.
TS26: inferior colliculus	Present	Regionally restricted	9b	The largely negative inferior colliculus/brachium of the inferior colliculus complex only appears labeled at its ventricular zone.
TS26: periaqueductal grey	Absent		9b

Structure	Level	Pattern	Image	Note
TS26: superior colliculus	Present	Regionally restricted	9c	Labeling was restricted to intermediate and superficial cell layers, plus the ventricular zone.
TS26: midbrain	Present	Regionally restricted	9c	Expression was detected in the preisthmus. Only part of the ependymal ventricular zone of the posterior intraencephalic sulcus expresses Otx2.
TS26: inferior colliculus	Present	Regionally restricted	9c	The inferior colliculus core is negative and appears surrounded deeply and superficially by a dispersed shell of positive cells.
TS26: midbrain-hindbrain junction	Present	Regionally restricted	9c	A number of positive preisthmic cells disperse in the neighboring isthmic mantle.
TS26: periaqueductal grey	Absent		9c

Structure	Level	Pattern	Image	Note
TS26: superior colliculus	Present	Regionally restricted	9d	Labeling was restricted to intermediate and superficial cell layers, plus the ventricular zone.
TS26: midbrain	Present	Regionally restricted	9d	Expression was detected in the preisthmus. Only part of the ependymal ventricular zone of the posterior intraencephalic sulcus expresses Otx2.
TS26: inferior colliculus	Present	Regionally restricted	9d	The inferior colliculus core is negative and appears surrounded deeply and superficially by a dispersed shell of positive cells.
TS26: midbrain-hindbrain junction	Present	Regionally restricted	9d	A number of positive preisthmic cells disperse in the neighboring isthmic mantle.
TS26: periaqueductal grey	Absent		9d