hGLP1R/hGIPR/hGCGR

Fig.1 Body weight change of HFD-induced hGLP1R/hGIPR/hGCGR obese mice treated with retatrutide. In high-fat diet (HFD)-induced hGLP1R/hGIPR/hGCGR obese mice, retatrutide significantly decreased body weight. The hGLP1R/hGIPR/hGCGR mice were fed CD or HFD. Saline or retatrutide was administered subcutaneously once daily until euthanasia. (A) Body weight and (B) body weight change were recorded daily (n=5-7, Mean ± SEM, ***p<0.001, Group2 vs. Group3).

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.2 Food intake of HFD-induced hGLP1R/hGIPR/hGCGR obese mice treated with retatrutide. In HFD-fed hGLP1R/hGIPR/hGCGR mice treated with retatrutide, food intake decreased during the first 8 days of dosing, but showed no significant change thereafter. The hGLP1R/hGIPR/hGCGR mice were fed CD or HFD. Saline or retatrutide was administered subcutaneously once daily until euthanasia. Food intake was recorded (n=5-7, Mean ± SEM,  ***p<0.001, Group2 vs. Group3). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.3 Fat mass and lean mass change of HFD-induced hGLP1R/hGIPR/hGCGR obese mice treated with retatrutide. In HFD-fed hGLP1R/hGIPR/hGCGR mice treated with retatrutide, both fat mass and lean mass were reduced. The hGLP1R/hGIPR/hGCGR mice were fed CD or HFD. Saline or retatrutide was administered subcutaneously once daily until euthanasia. (A) Fat mass, (B) fat mass change, (C) lean mass and (D) lean mass change were recorded  weekly (n=5-7, Mean ± SEM,  *p<0.05, ***p<0.001, Group2 vs. Group3). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.4 Blood glucose levels of HFD-induced hGLP1R/hGIPR/hGCGR obese mice treated with retatrutide. In high-fat diet (HFD)-induced hGLP1R/hGIPR/hGCGR obese mice, retatrutide significantly decreased blood glucose and improved glucose tolerance. The hGLP1R/hGIPR/hGCGR mice were fed CD or HFD. Saline or retatrutide was administered subcutaneously once daily until euthanasia. (A) An intraperitoneal glucose tolerance test (IPGTT) was performed by intraperitoneally injecting glucose and measuring blood glucose levels at 0, 15, 30, 60, 90 and 120 min post-injection( *p<0.05, **p<0.01, ***p<0.001, Group2 vs. Group3) and (B) the area under the curve (AUC) was calculated to evaluate systemic glucose tolerance. (n=5-7, Mean ± SEM,  *p<0.05, **p<0.01, ***p<0.001). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.5 Random blood glucose and insulin levels of HFD-induced hGLP1R/hGIPR/hGCGR obese mice treated with retatrutide. The hGLP1R/hGIPR/hGCGR mice were fed CD or HFD. Saline or retatrutide was administered subcutaneously once daily until euthanasia.  (A) Random blood glucose was measured weekly, and serum insulin levels were determined at the end of the study (n=5-7, Mean ± SEM,  ***p<0.001, Group2 vs. Group3). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.6 Serum lipid profiles of HFD-induced hGLP1R/hGIPR/hGCGR obese mice treated with retatrutide. In high-fat diet (HFD)-induced hGLP1R/hGIPR/hGCGR obese mice, retatrutide significantly decreased  serum total cholesterol and HDL-C levels. The hGLP1R/hGIPR/hGCGR mice were fed CD or HFD. Saline or retatrutide was administered subcutaneously once daily until euthanasia. Serum samples were collected to measure levels of (A) total cholesterol (T-CHO), (B) triglycerides (TG), (C) high-density lipoprotein cholesterol (HDL-C) and (D) low-density lipoprotein cholesterol (LDL-C) (n=5-7, Mean ± SEM,  ***p<0.001). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.7 Liver lipid profiles of HFD-induced hGLP1R/hGIPR/hGCGR obese mice treated with retatrutide. In high-fat diet (HFD)-induced hGLP1R/hGIPR/hGCGR obese mice, retatrutide significantly decreased liver triglycerides level. The hGLP1R/hGIPR/hGCGR mice were fed CD or HFD. Saline or retatrutide was administered subcutaneously once daily until euthanasia. Liver tissues were homogenized, and liver (A) triglycerides (TG) and (B) total cholesterol (T-CHO) levels were determined (n=5-7, Mean ± SEM,  *p<0.05, ***p<0.01). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.8 Liver weight of HFD-induced hGLP1R/hGIPR/hGCGR obese mice treated with retatrutide. In high-fat diet (HFD)-induced hGLP1R/hGIPR/hGCGR obese mice, retatrutide significantly decreased liver weight. The hGLP1R/hGIPR/hGCGR mice were fed CD or HFD. Saline or retatrutide was administered subcutaneously once daily until euthanasia. (A) Liver weight was recorded, and (B) the liver-to-body weight ratio (liver weight/BW) was calculated (n=5-7, Mean ± SEM,  *p<0.05, **p<0.01). 

Abbr: CD, chow diet. HFD, high-fat diet. BW, body weight.

Fig.9 White adipose tissue weight in hGLP1R/hGIPR/hGCGR obese mice after treatment with retatrutide. Retatrutide selectively reduced white adipose tissue mass, particularly in inguinal (iWAT), epididymal (eWAT) and mesenteric (mWAT) depots in hGLP1R/hGIPR/hGCGR triple-humanized obese mice. The hGLP1R/hGIPR/hGCGR mice were fed CD or HFD. Saline or retatrutide was administered subcutaneously once daily until euthanasia. (A) Inguinal (iWAT), (B) epididymal (eWAT), and (C) mesenteric (mWAT) white adipose tissues were dissected, weighed and normalized to body weight (BW) (D-F) (n=5-7, Mean ± SEM, *p<0.05, **p<0.01, ***p<0.001). 

Abbr: CD, chow diet. HFD, high-fat diet. BW, body weight.

Fig.10 Brown adipose tissue weight in hGLP1R/hGIPR/hGCGR obese mice after treatment with retatrutide. The hGLP1R/hGIPR/hGCGR mice were fed CD or HFD. Saline or retatrutide was administered subcutaneously once daily until euthanasia.  (A) Brown adipose tissue (BAT) was dissected, weighed, and (B) normalized to body weight (BW) (n=5-7, Mean ± SEM). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.11 Muscle weights of HFD-induced hGLP1R/hGIPR/hGCGR obese mice treated with retatrutide. In high-fat diet (HFD)-induced hGLP1R/hGIPR/hGCGR obese mice, administration of retatrutide significantly increased the tibialis anterior and gastrocnemius weight-to-body weight ratios relative to the saline-treated HFD group. The hGLP1R/hGIPR/hGCGR mice were fed CD or HFD. Saline or retatrutide was administered subcutaneously once daily until euthanasia.  (A-C) Soleus, tibialis anterior and gastrocnemius muscles were dissected and weighed, and each muscle weight was expressed relative to body weight (BW) (D-F) (n=5-7, Mean ± SEM,  *p<0.05, **p<0.01). 

Abbr: CD, chow diet. HFD, high-fat diet.