Probe:	mTcf-4 probe2
Probe preparation:	Antisense labelled with digoxigenin RNA
Visualized with:	Alkaline phosphatase
Assay notes:	Embryos were post-fixed in 0.2% glutaraldehyde/4% paraformaldehyde.

Structure	Level	Pattern	Image	Note
TS11: embryo endoderm	Present	Regionally restricted	4A	Expression is at the anterior endoderm.

Structure	Level	Pattern	Image	Note
TS10: embryo endoderm	Weak	Regionally restricted	4B	Expression is at the most anterior endoderm.

Structure	Level	Pattern	Image	Note
TS17: diencephalon	Strong		4C

Structure	Level	Pattern	Image	Note
TS13: rhombomere 5	Strong		4E
TS13: rhombomere 1	Present		4E
TS13: rhombomere 3	Present		4E
TS13: future spinal cord neural plate	Present	Regionally restricted	4E	Expression is at the most rostral border of the neural plate.

Structure	Level	Pattern	Image	Note
TS21: midbrain marginal layer	Present		4F
TS21: thalamus	Present		4F

Structure	Level	Pattern	Image	Note
TS19: diencephalon	Strong		4G
TS19: fronto-nasal process	Present		4G
TS19: limb	Present		4G
TS19: midbrain	Present	Scattered	4G	Expression is in a small number of scattered cells in the anteriormost part and lateral sides.

Structure	Level	Pattern	Image	Note
TS19: fronto-nasal process	Present		4H	Expression in these tissues is the same as in wild-type embryos.
TS19: limb	Present		4H	Expression in these tissues is the same as in wild-type embryos.
TS19: diencephalon	Absent		4H

Structure	Level	Pattern	Image	Note
TS15: forelimb bud	Absent		5A

Structure	Level	Pattern	Image	Note
TS19: forelimb bud	Present	Regionally restricted	5B	Expression is at the distal end and the proximal region.
TS19: hindlimb bud	Present	Regionally restricted	5B	Expression is at the proximal region.

Structure	Level	Pattern	Image	Note
TS20: digit mesenchyme	Present	Regionally restricted	5C	Expression is in the area of tarsal/carpal condensations.

Structure	Level	Pattern	Image	Note
TS21: forelimb phalanx pre-cartilage condensation	Present		5D
TS21: metacarpus pre-cartilage condensation	Present		5D
TS21: forelimb interdigital region	Absent		5D

Structure	Level	Pattern	Image	Note
TS21: limb mesenchyme	Present	Regionally restricted	5E	Expression is in mesenchymal cells surrounding the differentiating cartilage.
TS21: limb digit	Present	Regionally restricted	5E	Expression is in bones at the distal end of the digits.

Structure	Level	Pattern	Image	Note
TS15: future forebrain	Strong
TS15: future hindbrain	Absent

Structure	Level	Pattern	Image	Note
TS17: forelimb bud	Present	Regionally restricted		Expression is at the tip of the distal region.

Structure	Level	Pattern	Image	Note
TS17: diencephalon	Strong

Structure	Level	Pattern	Image	Note
TS17: diencephalon	Absent