hCB1(2)

Fig.1 Detection of CNR1 expression in cerebellum by RT-PCR. Cerebellum RNA was extracted from wild-type C57BL/6 mice and homozygous hCB1(2) knockin mice, then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human mRNA primers (n=2, male, 7 weeks old). Mouse Cnr1 mRNA (373 bp) was detectable only in wild-type C57BL/6 mice. Human CNR1 mRNA (209 bp) was detectable only in homozygous hCB1(2) knockin mice but not in wild-type mice. 

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

Fig.2 Detection of CNR1 expression in muscle, heart, kidney, epididymal White Adipose Tissue (eWAT of male) and gonadal White Adipose Tissue (gWAT of female) by RT-PCR. Mouse Cnr1 mRNA (373 bp) was detectable only in wild-type C57BL/6 mice. Human CNR1 mRNA (209 bp) was detectable only in homozygous hCB1(2) knockin mice but not in wild-type mice. Tissues RNA was extracted from 7-week-old male and female wild-type C57BL/6 (WT)(n=4) and homozygous hCB1(2) knockin mice (HO) (n=4), 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 CB1 expression in hCB1(2) mice by WB. CB1 protein was detectable obviously in brain and BAT, while not obvious in total tissue lysates of liver, kidney and mammary gland from both WT C57BL/6 and HO hCB1(2) mice, as the antibody is cross-reactived between human and mouse CB1. The brain, liver, kidney, mammary gland and BAT tissue lysates were collected from female 7-week-old wild-type C57BL/6 mice and homozygous hCB1(2) mice, and then analyzed by western blot with anti-CB1 antibody. 

Abbr. HO, homozygous; WT, wild type. M, marker; BAT, brown adipocyte tissue.

Note. The arrow indicates the target strip.

Fig.4 Detection of CB1 expression in hCB1(2) mice by WB. CB1 was detectable in eWAT/gWAT, iWAT, muscle, kidney and heart from both WT C57BL/6 and HO hCB1(2) mice , as the antibody was cross-reactived between human and mouse CB1. The eWAT/gWAT, iWAT, muscle, kidney and heart tissue lysates were collected from 6-7-week-old wild-type C57BL/6 mice and 7-week-old homozygous hCB1(2) mice, and then analyzed by western blot with anti-CB1 antibody. 

Abbr. HO, homozygous; WT, wild type; M, marker; WAT, white adipocyte tissue; PC, mouse brain tissue lysates.