hUGT1A1

Fig.1 Detection of UGT1A1 expression in liver by RT-PCR. Mouse Ugt1a1 mRNA (286 bp) was detectable both in wild-type C57BL/6 mice and heterozygous hUGT1A1 knockin mice. Human UGT1A1 mRNA (426 bp) was detectable only in heterozygous hUGT1A1 knockin mice but not in WT mice. Liver RNA was extracted from 12-week-old male WT C57BL/6 (n=2) and 11-week-old male heterozygous hUGT1A1 knockin mice (n=2), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers. 

Abbr. M, marker; HE, heterozygous; WT, wild type.

Fig.2 Detection of UGT1A1 expression in hUGT1A1 mice by WB. Human UGT1A1 was exclusively detectable in liver, small intestine and colon tissues from heterozygous hUGT1A1 mice with a human UGT1A1 specific antibody but not in WT mice. The liver, small intestine and colon tissue lysates were collected from 12-week-old male WT C57BL/6 mice and 11-week-old male heterozygous hUGT1A1 mice, and then analyzed by western blot. 

Abbr. HE, heterozygous; WT, wild type.

Fig.3 Blood Biochemical test of WT mice and hUGT1A1 mice. hUGT1A1 mice exhibited higher DBIL levels but similar TBIL, ALT, and AST levels compared to WT mice, indicating normal human UGT1A1 activity in catalyzing the glucuronidation of unconjugated bilirubin (UCB) to DBIL. Serum was collected from 4.5-month-old female wild-type C57BL/6 mice (n=5) and 4.5-month-old female heterozygous hUGT1A1 mice (n=4) to measure levels of (A) direct bilirubin (DBIL), (B) total brilirubin (TBIL), (C) alanine aminotransferase (ALT), and (D) aspartate aminotransferase (AST) (Mean±SEM, T-test, *p<0.05; ns, no significance). 

Abbr. HE, heterozygous; WT, wild type.