Antibody:	anti-Nkx-2.5 (N-19): sc-8697
Detection system:	Secondary antibody coupled to horseradish peroxidase

Structure	Level	Pattern	Image	Note
TS15: foregut epithelium	Present	Regionally restricted	1b	Expressed in the ventral endoderm.
TS15: outflow tract mesenchyme	Present		1b
TS15: visceral pericardium	Present	Regionally restricted	1b	Expressed in the coelomic wall bordering the outflow tract. Expression was more pronounced at the left side of the outflow tract.

Structure	Level	Pattern	Image	Note
TS17: heart	Strong	Regionally restricted	1f	Expressed in the anterior heart field, including the coelomic wall, and in the outflow tract myocardium.
TS17: outflow tract cardiac muscle	Strong	Clusters	1f	Expressed as a central cluster of cells ventral to the foregut, extending preferentially on the left side of the aortic sac and surrounding the left 6th pharyngeal arch artery. Absent on the right side in the region bordering the aortic wall.
TS17: outflow tract endocardial tube	Strong	Regionally restricted	1f	Expressed in the left-side region of the coelomic wall. Largely absent expression in the coelomic wall on the right side.
TS17: cardiac muscle tissue	Present		1f

Structure	Level	Pattern	Image	Note
TS19: outflow tract	Present	Regionally restricted	2c	Expressed as a mesenchymal column lining the pulmonary wall on the left side of the outflow tract and merging with the subpulmonary myocardium. There is no such expression on the aortic side.
TS19: cardiac muscle tissue	Present		2c

Structure	Level	Pattern	Image	Note
TS19: outflow tract	Present	Regionally restricted	2f	Expressed in a column of myocardial precursor cells exclusively on the left side, where the pulmonary trunk and right ventricular outflow tract are developing.
TS19: left atrium cardiac muscle	Present		2f
TS19: left ventricle cardiac muscle	Present		2f
TS19: right atrium cardiac muscle	Present		2f
TS19: right ventricle cardiac muscle	Present		2f

Structure	Level	Pattern	Image	Note
TS17: outflow tract cardiac muscle	Present	Regionally restricted	4b	Expressed as an elongated column extending along the left side.
TS17: heart mesentery	Present	Regionally restricted	4b	Expressed as a well-organized cluster of cells in the cardiac mesenchyme.
TS17: pharynx mesenchyme	Present	Regionally restricted	4b	Expressed as a group of positive cells located behind the heart in the pre-pharyngeal mesoderm.

Structure	Level	Pattern	Image	Note
TS17: heart mesentery	Present	Regionally restricted	4e	Expressed as in wild-type in a cluster of cells in the cardiac mesenchyme.
TS17: outflow tract cardiac muscle	Present	Regionally restricted	4e	Expressed in an abnormal very short column of cells extending along the left side.

Structure	Level	Pattern	Image	Note
TS19: pharynx mesenchyme	Present	Regionally restricted		Expressed as a central cluster of cells located in the pre-pharyngeal mesoderm dorsal to the level of the outflow tract.

Structure	Level	Pattern	Image	Note
TS20: outflow tract	Present	Regionally restricted		Expressed in a column of mesenchymal cells found exclusively within the indentation at the orifice level at the pulmonary side but not in the aortic region.

Structure	Level	Pattern	Image	Note
TS22: outflow tract	Absent			The mesenchymal column of expressing cells was no longer present.