hMTARC1

Fig.1 Detection of MTARC1 expression in liver by RT-PCR. 

Wild type: only one band at 211 bp with primers F1/R1(mMtarc1);

Homozygous: only one band at 211 bp with primers F2/R2(hMTARC1). 

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

Fig.2 Detection of human MTARC1 expression in hMTARC1 Mice by WB. 

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

Note. The anti-human MTARC1 antibody cross-reacted with mouse MTARC1 and humanized MTARC1.

Fig.3 Detection of MTARC1 expression in liver by qPCR. Mouse Mtarc1 mRNA was detectable only in wild-type mice but not in homozygous hMTARC1 mice. Human MTARC1 mRNA was detectable only in homozygous hMTARC1 mice but not in wild-type mice. Liver RNA were extracted from 8-16-week-old male wild-type C57BL/6 (n=5 for each age) and homozygous hMTARC1 mice (n=5 for each age), then cDNA libraries were synthesized by reverse transcription, followed by qPCR with human MTARC1, mouse Mtarc1 and mouse Gapdh primers. (A) Relative expression represents the human MTARC1 mRNA level relative to its expression in 8-week-old male hMTARC1 mice. (B) Relative expression represents the mouse Mtarc1 mRNA level relative to its expression in 8-week-old male wild-type mice.

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

Fig.4 Detection of human and mouse MTARC1 protein levels in liver of WT C57BL/6 mice and hMTARC1 mice by ELISA. MTARC1 protein was detectable in liver from both WT C57BL/6 and hMTARC1 mice, as both of the ELISA kits were cross-reactived between human and mouse MTARC1 protein. Lysates of liver tissues were collected from wild-type C57BL/6 mice (n=5 for each age) and homozygous hMTARC1 mice (n=5 for each age), and analyzed by ELISA with anti-human MTARC1 ELISA kit (Fine-test, EH4968) or anti-mouse MTARC1 ELISA kit. 

Abbr.  HO, homozygous; WT, wild type.

Fig.5 Liver function indicators of hMTARC1 mice. hMTARC1 mice exhibited serum ALT and AST levels similar to those of WT mice. Serum samples were collected from WT C57BL/6 mice and homozygous hMTARC1 mice to measure levels of (A) alanine aminotransferase (ALT), and (B) aspartate aminotransferase (AST) (n=5, Mean ± SEM, Unpaired t-test, *p＜0.05). 

Abbr. WT, wild-type; HO, homozygous; WD, Western diet.

Fig.6 Blood lipid profiles of hMTARC1 mice. hMTARC1 mice exhibited serum TG, T-CHO, HDL-C and LDL-C levels similar to those of WT mice. Serum samples were collected from WT C57BL/6 mice and homozygous hMTARC1 mice to measure levels of (A) triglycerides (TG), (B) total cholesterol (T-CHO), (C) high-density lipoprotein cholesterol (HDL-C) and (D) low-density lipoprotein cholesterol (LDL-C) (n=5, Mean ± SEM, Unpaired t-test, *p＜0.05; **p＜0.01; ***p＜0.0001). 

Abbr. WT, wild-type; HO, homozygous; WD, Western diet.