hCB1

Fig1. Detection of CB1 expression in cerebellum by RT-PCR. 

Wild type: only one band at 373 bp with primers F1/R1(mCb1);

Homozygous: only one band at 209 bp with primers F2/R2(hCB1);

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

Fig2. Detection of human CB1 expression in Brains, liver and kidney in hCB1 knockin mice by WB. 

Abbr. HO, homozygous; WT, wild type.

Note. The anti-human CB1 Antibody cross-reacted with mouse CB1 and humanized CB1.

Fig3. Detection of human CB1 expression in brain (Hippocampus) in hCB1 knockin mice by IF. 

Brains were collected from wild-type mice and HO hCB1 mice, and analyzed by IF with anti-human CB1 antibody. 

Abbr. HO, homozygous; WT, wild type.

Note. The anti-human CB1 Antibody cross-reacted with mouse CB1 and humanized CB1.

Fig4. Detection of human CB1 expression in CA1 in hCB1 knockin mice by IF. 

Brains were collected from wild-type mice and HO hCB1 mice, and analyzed by IF with anti-human CB1 antibody. 

Abbr. HO, homozygous; WT, wild type. 

Note. The anti-human CB1 Antibody cross-reacted with mouse CB1 and humanized CB1.

Fig5. Detection of human CB1 expression in CA3 in hCB1 knockin mice by IF. 

Brains were collected from wild-type mice and HO hCB1 mice, and analyzed by IF with anti-human CB1 antibody. 

Abbr. HO, homozygous; WT, wild type. 

Note. The anti-human CB1 Antibody cross-reacted with mouse CB1 and humanized CB1.

Fig6. Detection of human CB1 expression in dentate gyrus(DG) in hCB1 knockin mice by IF. 

Brains were collected from wild-type mice and HO hCB1 mice, and analyzed by IF with anti-human CB1 antibody. 

Abbr. HO, homozygous; WT, wild type. 

Note. The anti-human CB1 Antibody cross-reacted with mouse CB1 and humanized CB1.

Fig7. Detection of human CB1 expression in cerebellum in hCB1 knockin mice by IF. 

Brains were collected from wild-type mice and HO hCB1 mice, and analyzed by IF with anti-human CB1 antibody. 

Abbr. HO, homozygous; WT, wild type. 

Note. The anti-human CB1 Antibody cross-reacted with mouse CB1 and humanized CB1.

Fig8. Detection of human CB1 mRNA knockdown in iWAT, pgWAT and BAT by qPCR. 8-week-old, male homozygous hCB1 (HO) mice were assigned to three groups (n=4 per group). Mice in Group 1 received a single subcutaneous injection of vehicle, while mice in Group 2 and 3 were administered a single subcutaneous dose of the test drug, a small nucleic drug from a client. At 28 days post-dosing, the inguinal (iWAT) and perigonadal (pgWAT) white adipose tissue and brown adipose tissue (BAT) were harvested to quantify human CB1 mRNA levels. Data were normalized to mouse Ppia mRNA levels. Results indicated that the baseline expression of hCB1 mRNA was lower in BAT relative to WAT. Notably, high-dose administration of the test drug elicited a significant downregulation (over 50%) of human CB1 mRNA levels especially within the WAT.