hKHK

Fig.1 Detection of KHK expression in liver, kidney and small intestine by RT-PCR. Mouse Khk mRNA (165 bp) was detectable only in wild-type C57BL/6 mice. Human KHK mRNA (183 bp) was detectable only in homozygous hKHK knockin mice but not in wild-type mice. Liver, kidney and small intestine RNA was extracted from 6-week-old female wild-type C57BL/6 (WT)(n=3) and homozygous hKHK knockin mice (HO) (n=3), 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 KHK expression in liver, kidney and small intestine by qPCR. Mouse Khk mRNA was detectable only in wild-type C57BL/6 mice. Human KHK mRNA was detectable only in homozygous hKHK knockin mice but not in wild-type mice. Liver, kidney and small intestine RNA were extracted from 6-week-old female wild-type C57BL/6 (WT)(n=3) and homozygous hKHK knockin mice (HO) (n=3), then cDNA libraries were synthesized by reverse transcription, followed by qPCR with human KHK, mouse Khk and mouse Gapdh primers. Relative expression represents the human KHK mRNA level relative to its average expression in homozygous hKHK knockin mice. Relative expression represents the mouse Khk mRNA level relative to its average expression in wild-type mice.

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

Fig.3 Detection of KHK expression in hKHK mice by WB. KHK was detectable in liver and kidney from both WT C57BL/6 and HO hKHK mice , as the antibody was cross-reactived between human and mouse KHK. The liver and kidney tissue lysates were collected from 7-week-old male wild-type C57BL/6 mice and 7-week-old male homozygous hKHK mice, and then analyzed by western blot with anti-KHK antibody. 

Abbr. HO, homozygous; WT, wild type; PC, positive control, HepG2 cells.