hZNF423(CDS)

Fig.1 Detection of ZNF423 expression in subcutaneous fat pad by RT-PCR. Mouse Zfp423 mRNA (715 bp) was detectable only in wild-type C57BL/6 mice. Human ZNF423 mRNA (577 bp) was detectable only in homozygous hZNF423(CDS) knock-in mice but not in wild-type mice. Subcutaneous fat pad RNA was extracted from 7-week-old female wild-type C57BL/6 (n=2) and homozygous hZNF423(CDS) knock-in mice (n=2), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers. 

Abbr. M, marker; HO, homozygous; WT, wild type.

Fig.2 Detection of ZNF423 expression in bone marrow and subcutaneous fat pad by RT-PCR. Mouse Zfp423 mRNA (205 bp) was detectable only in wild-type C57BL/6 mice. Human ZNF423 mRNA (577 bp) was detectable only in homozygous hZNF423(CDS) knock-in mice but not in wild-type mice. Bone marrow and subcutaneous fat pad RNA was extracted from 7-week-old female wild-type C57BL/6 (WT)(n=2) and homozygous hZNF423(CDS) knock-in mice (HO) (n=2), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers. 

Abbr. M, marker; HO, homozygous; WT, wild type.

Fig.3 Detection of ZNF423 expression in subcutaneous fat pad by qPCR. H/M Zfp423 mRNA was detectable both in wild-type mice and homozygous hZNF423(CDS) knock-in mice. Subcutaneous fat pad RNA was extracted from 7-week-old female wild-type C57BL/6 (n=2) and homozygous hZNF423(CDS) knock-in mice (n=2), then cDNA libraries were synthesized by reverse transcription, followed by qPCR with h/m Zfp423 and mouse Gapdh primers. Relative expression represents the h/m Zfp423 mRNA level relative to its average expression in wild-type mice . 

Abbr. M, marker; HO, homozygous; WT, wild type.

Fig.4 Detection of human ZNF423 expression in hZNF423 (CDS) knock-in mice by IHC. ZNF423 protein was detectable in the thalamic nucleus of brain from WT mice and hZNF423 (CDS) knock-in mice, as the antibody was cross-actived between human and mouse ZNF423 protein. Brain tissues were collected from 14-week-old male WT C57BL/6 mice and 14 or 16-week-old male homozygous hZNF423(CDS) mice, and then analyzed by IHC with anti-ZNF423 antibody. Black arrows indicate positive staining (brown). The scales are same in each image. Magnification, 20X.

Abbr. HO, homozygous; WT, wild type.

Fig.5 Detection of human ZNF423 expression in hZNF423 (CDS) knock-in mice by IHC. ZNF423 protein was detectable in the cerebellum of brain from WT mice and hZNF423 (CDS) knock-in mice, as the antibody was cross-actived between human and mouse ZNF423 protein. Brain tissues were collected from 14-week-old male WT C57BL/6 mice and 14 or 16-week-old male homozygous hZNF423(CDS) mice, and then analyzed by IHC with anti-ZNF423 antibody. Black arrows indicate positive staining (brown). The scales are same in each image. Magnification, 20X.

Abbr. HO, homozygous; WT, wild type.