Antibody:	Phf6 antibody
Detection system:	Secondary antibody coupled to Alexa Fluor
Assay notes:	Freshly frozen and freshly cut embryos or brains were fixed first in methanol and then in 2% paraformaldehyde. The AlexaFluor 568 (red) or 488 (green) labeled secondary antibodies were used.

Structure	Level	Pattern	Image	Note
TS28: testis	Present		5I

Structure	Level	Pattern	Image	Note
TS20: cerebral cortex	Present		6A

Structure	Level	Pattern	Image	Note
TS22: cerebral cortex ventricular layer	Strong		6B

Structure	Level	Pattern	Image	Note
TS24: cerebral cortex	Strong		6C

Structure	Level	Pattern	Image	Note
TS26: cerebral cortex	Moderate	Regionally restricted	6D	Expression is in post-mitotic neurons.

Structure	Level	Pattern	Image	Note
TS27: cerebral cortex	Strong	Regionally restricted	6E	Expression is in post-mitotic neurons.

Structure	Level	Pattern	Image	Note
TS28: cerebral cortex	Weak		6F

Structure	Level	Pattern	Image	Note
TS27: hippocampus granule cell layer	Strong	Regionally restricted	6G	Expression is in post-mitotic neurons.

Structure	Level	Pattern	Image	Note
TS27: cerebellum Purkinje cell layer	Strong	Regionally restricted	6H	Expression is in post-mitotic neurons.
TS27: cerebellum internal granule cell layer	Strong	Regionally restricted	6H	Expression is in post-mitotic neurons.

Structure	Level	Pattern	Image	Note
TS28: cerebral cortex	Weak		6I

Structure	Level	Pattern	Image	Note
TS17: central nervous system	Strong			Nuclear expression is moderate in proliferating cells and is strong in post-mitotic neurons.
TS17: future adenohypophysis	Strong	Regionally restricted		Nuclear expression is in the anterior pituitary.

Structure	Level	Pattern	Image	Note
TS20: central nervous system	Strong			Nuclear expression is moderate in proliferating cells and is strong in post-mitotic neurons.
TS20: adenohypophysis	Moderate
TS20: limb	Present
TS20: liver	Present
TS20: nasal process	Present
TS20: peripheral nervous system	Present
TS20: epidermis	Weak
TS20: intestine epithelium	Weak
TS20: neurohypophysis	Weak
TS20: blood	Absent
TS20: cardiac muscle tissue	Absent
TS20: skeletal muscle	Absent

Structure	Level	Pattern	Image	Note
TS22: central nervous system	Moderate			Nuclear expression is weak in proliferating cells and is moderate in post-mitotic neurons.
TS22: adenohypophysis	Present
TS22: liver	Trace	Single cells		Some hepatocytes have weak nuclear expression and some have no expression.
TS22: blood	Absent
TS22: cardiac muscle tissue	Absent
TS22: cartilage	Absent
TS22: intestine epithelium	Absent
TS22: lung epithelium	Absent
TS22: lung mesenchyme	Absent
TS22: neurohypophysis	Absent
TS22: skeletal muscle	Absent
TS22: nose	Absent			Absent in the nasal process.
TS22: metanephros	Absent			Absent in the epithelial cells and mesenchyme.

Structure	Level	Pattern	Image	Note
TS24: central nervous system	Moderate			Nuclear expression is weak in proliferating cells and is moderate in post-mitotic neurons.
TS24: adenohypophysis	Present
TS24: olfactory epithelium	Present
TS24: epidermis	Weak			Some cells have weak nuclear expression and some have no expression.
TS24: lung epithelium	Weak			Some cells have weak nuclear expression and some have no expression.
TS24: liver	Weak			Some hepatocytes have weak nuclear expression and some have no expression.
TS24: metanephros	Weak			Some epithelial cells have weak nuclear expression, absent in the mesenchyme.
TS24: blood	Absent
TS24: cardiac muscle tissue	Absent
TS24: cartilage	Absent
TS24: intestine epithelium	Absent
TS24: intestine mesenchyme	Absent
TS24: lung mesenchyme	Absent
TS24: neurohypophysis	Absent
TS24: skeletal muscle	Absent
TS24: tongue muscle	Absent
TS24: dermis	Absent			Absent in hair follicle cells.
TS24: nose	Present	Regionally restricted		Absent in nasal mesenchyme, present in olfactory epithelium.

Structure	Level	Pattern	Image	Note
TS26: central nervous system	Present			Nuclear expression is in post-mitotic neurons and is weak in proliferating cells.
TS26: adenohypophysis	Present
TS26: peripheral nervous system	Weak	Regionally restricted		Nuclear expression is in post-mitotic neurons.
TS26: olfactory epithelium	Weak
TS26: tongue papillae	Weak	Regionally restricted		Nuclear expression is in taste bud cells.
TS26: thymus	Weak	Regionally restricted		Nuclear expression is in the epithelium, absent in thymocytes.
TS26: enteric nervous system	Weak	Regionally restricted		Nuclear expression is in the enteric neurons.
TS26: gut	Weak	Regionally restricted		Nuclear expression is in the enteric neurons.
TS26: liver	Weak	Regionally restricted		Some hepatocytes have weak nuclear expression and some have no expression.
TS26: nose	Weak	Regionally restricted		Absent in nasal mesenchyme, present in olfactory epithelium.
TS26: blood	Absent
TS26: cardiac muscle tissue	Absent
TS26: cartilage	Absent
TS26: dermis	Absent
TS26: epidermis	Absent
TS26: intestine epithelium	Absent
TS26: skeletal muscle	Absent
TS26: tongue muscle	Absent

Structure	Level	Pattern	Image	Note
TS28: central nervous system	Present	Regionally restricted		Nuclear expression is in post-mitotic neurons.
TS28: thymus	Weak	Regionally restricted		Nuclear expression is in the epithelium, absent in thymocytes.
TS28: cardiac muscle tissue	Absent
TS28: dermis	Absent
TS28: epidermis	Absent
TS28: lung connective tissue	Absent
TS28: lung epithelium	Absent
TS28: skeletal muscle	Absent
TS28: liver	Absent			Absent in hepatocytes.
TS28: intestine	Absent			Absent in epithelial cells and mesenchyme.

Structure	Level	Pattern	Image	Note
TS28: brain	Weak			Protein was detectable at low levels in all neurons.

Structure	Level	Pattern	Image	Note
TS28: main olfactory bulb mitral cell layer	Strong		7A	Expression is nuclear.
TS28: main olfactory bulb	Weak	Regionally restricted	7A	Expression is in olfactory bulb interneurons, i.e. granule cells and periglomerular cells.

Structure	Level	Pattern	Image	Note
TS28: main olfactory bulb mitral cell layer	Strong		7B	Expression is nuclear.
TS28: main olfactory bulb	Weak	Regionally restricted	7B	Expression is in olfactory bulb interneurons, i.e. granule cells and periglomerular cells.

Structure	Level	Pattern	Image	Note
TS28: main olfactory bulb mitral cell layer	Strong		7C	Expression is nuclear.

Structure	Level	Pattern	Image	Note
TS28: cerebellum Purkinje cell layer	Strong		7D	Expression is nuclear.
TS28: cerebellum granule cell layer	Weak		7D

Structure	Level	Pattern	Image	Note
TS28: cerebellum Purkinje cell layer	Strong		7E	Expression is nuclear and is low or absent in astrocytes.

Structure	Level	Pattern	Image	Note
TS28: cerebellum Purkinje cell layer	Strong		7F	Expression is nuclear and is low or absent in astrocytes.

Structure	Level	Pattern	Image	Note
TS28: cortical layer V	Present		7H	Expression is nuclear.

Structure	Level	Pattern	Image	Note
TS28: cortical layer V	Present		7I	Expression is nuclear.

Structure	Level	Pattern	Image	Note
TS17: brain	Present	Regionally restricted	7J	Expression is in proliferating neural precursors.

Structure	Level	Pattern	Image	Note
TS27: brain subventricular zone	Present		7K

Structure	Level	Pattern	Image	Note
TS27: hippocampus granule cell layer	Present		7L

Structure	Level	Pattern	Image	Note
TS27: brain subventricular zone	Present		7N

Structure	Level	Pattern	Image	Note
TS27: brain subventricular zone	Present		7O

Structure	Level	Pattern	Image	Note
TS17: future adenohypophysis	Strong		8C , 8B	Expression is nuclear.

Structure	Level	Pattern	Image	Note
TS20: adenohypophysis	Strong		8F , 8E	Expression is nuclear.
TS20: neurohypophysis	Moderate		8F , 8E

Structure	Level	Pattern	Image	Note
TS22: adenohypophysis	Moderate		8H , 8I
TS22: neurohypophysis	Absent		8H , 8I	Very low or no levels.

Structure	Level	Pattern	Image	Note
TS24: adenohypophysis	Moderate		8L , 8K
TS24: neurohypophysis	Absent		8K , 8L	Very low or no levels.

Structure	Level	Pattern	Image	Note
TS20: nasal process	Strong		9C , 9B

Structure	Level	Pattern	Image	Note
TS20: limb mesenchyme	Strong		9F , 9E

Structure	Level	Pattern	Image	Note
TS28: spermatid	Present	Single cells	9H , 9I	Expression is in spermatocytes and round spermatids, but is low or not present in spermatogonia and elongated spermatids.
TS28: spermatocyte	Present	Single cells	9H , 9I	Expression is in spermatocytes and round spermatids, but is low or not present in spermatogonia and elongated spermatids.
TS28: spermatogonium	Trace	Single cells	9H , 9I	Expression is in spermatocytes and round spermatids, but is low or not present in spermatogonia and elongated spermatids.

Structure	Level	Pattern	Image	Note
TS28: spermatid	Present	Single cells	9J	Expression is in spermatocytes and round spermatids, but is low or not present in spermatogonia and elongated spermatids.
TS28: spermatocyte	Present	Single cells	9J	Expression is in spermatocytes and round spermatids, but is low or not present in spermatogonia and elongated spermatids.
TS28: spermatogonium	Trace	Single cells	9J	Expression is in spermatocytes and round spermatids, but is low or not present in spermatogonia and elongated spermatids.

Structure	Level	Pattern	Image	Note
TS28: spermatid	Present	Single cells	9K	Expression is in spermatocytes and round spermatids, but is low or not present in spermatogonia and elongated spermatids.
TS28: spermatocyte	Present	Single cells	9K	Expression is in spermatocytes and round spermatids, but is low or not present in spermatogonia and elongated spermatids.
TS28: spermatogonium	Trace	Single cells	9K	Expression is in spermatocytes and round spermatids, but is low or not present in spermatogonia and elongated spermatids.

Structure	Cell Type	Level	Pattern	Image	Note
TS28: Leydig cell	Leydig cell	Weak	Regionally restricted		Nuclear expression in Leydig cells.
TS28: Sertoli cell	Sertoli cell	Weak	Regionally restricted		Nuclear expression in Sertoli cells.
TS28: spermatid		Present	Single cells		Expression is in primary spermatocytes, secondary spermatocytes, and round spermatids, but is low or absent in spermatogonia, spermatozoa, and elongated spermatids.
TS28: spermatocyte		Present	Single cells		Expression is in primary spermatocytes, secondary spermatocytes, and round spermatids, but is low or absent in spermatogonia, spermatozoa, and elongated spermatids.
TS28: spermatogonium		Trace	Single cells		Expression is in primary spermatocytes, secondary spermatocytes, and round spermatids, but is low or absent in spermatogonia, spermatozoa, and elongated spermatids.
TS28: spermatozoon		Trace	Single cells		Expression is in primary spermatocytes, secondary spermatocytes, and round spermatids, but is low or absent in spermatogonia, spermatozoa, and elongated spermatids.