Gcgr-KO(2)

Fig.1 Detection of Gcgr expression in liver by RT-PCR. Mouse Gapdh mRNA (123 bp) was detectable both in wild-type C57BL/6 mice and homozygous Gcgr-KO(2) mice. Mouse Gcgr mRNA (166 bp) was detectable only in wild-type mice but not in homozygous Gcgr-KO(2) mice. Liver RNA was extracted from 8-week-old male and female wild-type C57BL/6 (WT) (n=2) and homozygous Gcgr-KO(2) 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.2 Detection of Gcgr expression in liver by qPCR. Mouse Gcgr mRNA was detectable only in wild-type mice but not in homozygous Gcgr-KO(2) mice. Liver RNA were extracted from 8-week-old male and female wild-type C57BL/6 (WT) (n=2) and homozygous Gcgr-KO(2) mice (HO) (n=2), then cDNA libraries were synthesized by reverse transcription, followed by qPCR with mouse Gcgr and mouse Gapdh primers. Relative expression represents the mouse Gcgr mRNA level relative to its average expression in wild-type mice. 

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

Fig.3 Body weight and tissues weight of WT C57BL/6 mice and Gcgr-KO(2) mice without fasting. Gcgr-KO(2) mice exhibited significant decreases in liver weight and kidney weight compared to WT C57BL/6 mice.

Abbr. HO, homozygous; WT, wild type; eWAT, epididymal white adipose tissue; sWAT, subcutaneous white adipose tissue.

Fig.4 Detection of glucagon levels in plasma of WT C57BL/6 mice and HO Gcgr-KO(2) mice by ELISA. In contrast to the low baseline glucagon in WT C57BL/6 mice, Gcgr-KO(2) mice developed severe compensatory hyperglucagonemia resulting from defective negative feedback. Plasma was collected from wild-type C57BL/6 mice (n=5, male, 18 weeks old) and HO Gcgr-KO(2) mice (n=4, male, 18 weeks old) without fasting, and analyzed by ELISA.

Abbr.  HO, homozygous; WT, wild type; N.D. not detected.

Fig.5 Detection of TG levels in liver of WT C57BL/6 mice and HO Gcgr-KO(2) mice by ELISA. Gcgr-KO(2) mice exhibited an obvious decrease in hepatic TG levels relative to WT C57BL/6 mice, possibly reflecting a decreased hepatic TG uptake in the Gcgr-KO mice. Liver homogenates were collected from wild-type C57BL/6 mice (n=5, male, 18 weeks old) and HO Gcgr-KO(2) mice (n=4, male, 18 weeks old) without fasting, and analyzed by ELISA.

Abbr.  HO, homozygous; WT, wild type.

Fig.6 Blood biochemical test of WT C57BL/6 mice and HO Gcgr-KO(2) mice. Gcgr-KO(2) mice exhibited significant lower ALT and AST levels and higher TG, LDL-C and NEFA levels compared to WT mice. Serum was collected from wild-type C57BL/6 mice (n=5, male, 18 weeks old) and HO Gcgr-KO(2) mice (n=4, male, 18 weeks old) without fasting to detecte blood biochemical profiles. 

Abbr.  HO, homozygous; WT, wild type.