B-hGIPR mice_Biocytogen Pharmaceuticals (Beijing) Co., Ltd._Biocytogen, drug development, antibody discovery

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동물/세포 모델

동물/세포모델

Humanized Mouse Models

GPCR Humanized Mice

Text

B-hGIPR mice

Strain Name	C57BL/6-Gipr^{tm2(GIPR)Bcgen}/Bcgen	Common Name	B-hGIPR mice
Background	C57BL/6	Catalog number	112714
Aliases	PGQTL2

mRNA expression analysis

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Strain specific analysis of GIPR mRNA expression in wild-type C57BL/6 mice and B-hGIPR mice by RT-PCR. Brain, eWAT and ingWAT RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hGIPR mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human GIPR primers. Mouse Gipr mRNA was detectable only in wild-type C57BL/6 mice. Human GIPR mRNA was detectable only in homozygous B-hGIPR mice but not in wild-type mice. eWAT, epididymal white adipose tissue; ingWAT, inguinal white adipose tissue.

GIPR expression analysis by WB

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Western blot analysis of GIPR protein expression in homozygous B-hGIPR mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hGIPR mice (H/H), and then analyzed by western blot with anti-GIPR antibody. 40 μg total proteins were loaded for western blotting analysis. GIPR was detected in brain, stomach, pancreas and liver.

GIPR expression analysis by IHC

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Representative GIPR expression in different tissues of B-hGIPR mice by IHC. Tissues were stained with antibodies that recognized with both human and mouse GIPR (B-J) or anti-IgG antibody (A). Pancreas and brain of B-hGIPR mice and wild-type C57BL/6 mice show GIPR positive. Liver and adipose of B-hGIPR mice and wild-type C57BL/6 mice show GIPR negative. Mouse tissues from the wild-type C57BL/6 mice (A-E). Human pancreas carcinoma as a positive control (F). Other tissues were got from homozygous B-hGIPR mice (G-J). Original magnification ×200. Abbreviations: IHC, immunohistochemistry.

In vivo function of exogenous GIP during IGPTT

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Analyzed by 2way-ANOVA,*P<0.05, **P<0.01, ***P<0.001

In vivo function of m/hGIP during IGPTT

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In vivo function of m/hGIP during IGPTT. Plasma glucose and insulin concentrations were measured after intraperitoneal injection of 50 nmol/kg mouse GIP (Cat. HY-P77948, MCE) or human GIP (Cat. HY-P0276, MCE) and glucose (40%) in wild-type C57BL/6J mice and B-hGIPR mice. (male, 6-week-old, n=6). Data represent means ± SEM. Analyzed by 2way-ANOVA,*P<0.05, **P<0.01, ***P<0.001

In vivo efficacy in HFD induced B-hGIPR mice

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Efficacy study in HFD induced B-hGIPR mice. B-hGIPR mice were fed with a high-fat diet for 12 weeks to induce mice obesity. (A) Body weight change after HFD induction. (B-C) Body weight change after hGIPR-antibody Amgen (2G10) analog (in house), Semaglutide, and combination treatment. (D) Effect of hGIPR-antibody Amgen (2G10), Semaglutide, and combination treatment on food intake on Day 21. 8 mice per group. Values are expressed as mean ± SEM. Significance was determined by the Ordinary one-way ANOVA compared with HFD PBS group. *p<0.05, **p<0.01,***p<0.001 ,****p<0.0001.

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Efficacy study in HFD induced B-hGIPR mice. (A) Fasting insulin after treatment in plasma after 6 hours of fasting. (B) After fasting for 6 hours, mice were intraperitoneally injected with 15% Glucose (1.5 g/kg) for Glucose Tolerance Tests after treatment. (C-F) Blood biochemical analysis after treatment. Values are expressed as mean ± SEM. Significance was determined by the Ordinary one-way ANOVA compared with HFD PBS group. *p<0.05, **p<0.01,***p<0.001 ,****p<0.0001.

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Efficacy study in HFD induced B-hGIPR mice. (A-D) Weight of white adipose tissue at the end of the treatment. Values are expressed as mean ± SEM. Significance was determined by the Ordinary one-way ANOVA compared with HFD PBS group. . *p<0.05, **p<0.01,***p<0.001 ,****p<0.0001.

In vivo function of hGIP and GIPR-antibody in HFD induced B-hGIPR mice

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In vivo function of hGIP and GIPR-antibody in HFD induced B-hGIPR mice. B-hGIPR mice were fed with a high-fat diet for 12 weeks to induce mice obesity and were randomized into groups and received vehicle or GIPR antibody Amgen (2G10) analog (in house). Approximately 24 hours later, saline or 0.25 mg/kg human GIP (Cat. HY-P0276, MCE) was given via IP injection, then immediately followed with an IP injection of glucose at 1.5 g/kg. Blood glucose and plasma insulin were measured during IGPTT . Data represent means ± SEM. Analyzed by 2 way-ANOVA, *P<0.05, **P<0.01, ***P<0.001.