Strain specific analysis of INHBE mRNA expression in wild-type C57BL/6 mice and B-hINHBE mice by RT-PCR. Liver RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hINHBE mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human INHBE primers. Mouse Inhbe mRNA was detectable only in wild-type C57BL/6 mice. Human INHBE mRNA was detectable only in homozygous B-hINHBE mice but not in wild-type mice. 

Western blot analysis of INHBE protein expression in homozygous B-hINHBE mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hINHBE mice (H/H), and then analyzed by western blot with anti-INHBE antibody. 40 μg total proteins were loaded for western blotting analysis. INHBE was detected in liver and kidney of wild-type mice and homozygous B-hINHBE mice, as the antibody cross-recognized both human and mouse INHBE.

Relative mRNA levels of hINHBE in B-hINHBE mice by qPCR. B-hINHBE mice were randomly divided into two groups (n=3/group, 8 weeks old, male).  B-hINHBE mice were fasted or fed 16 hours, and the liver tissue was collected to detect the expression level of human INHBE mRNA by qPCR. The expression level of INHBE in the fasted group of mice was significantly increased. Consistent with the function of INHBE in preventing lipolysis. Values are expressed as mean ± SEM. 

The inhibitory efficiency of the nucleic acid drugs against human INHBE in B-hINHBE mice. B-hINHBE mice were randomly divided into two groups (male, 9-10 weeks old). The human INHBE targeted nucleic acid drugs (provided by a client) and PBS were administered to the mice individually. The nucleic acid drugs was administered in the form of PBS aqueous solution. The mice were sacrificed on day 5, and the liver tissue was collected to detect the expression level of human INHBE mRNA by qPCR. (A) The schematic diagram of experimental processing. (B) The expression of human INHBE mRNA in the liver. The human INHBE in the CPD-1 group was significantly reduced compared to the control group. Values are expressed as mean ± SEM.