hGPR65

Fig.1 Detection of Gpr65 expression in spleen and lung by RT-PCR. Mouse Gpr65 mRNA (846 bp) was detectable both in wild-type C57BL/6 mice. Human GRB14 mRNA (259 bp) was detectable only in homozygous hGPR65 knock-in mice but not in wild-type mice. Spleen and lung RNA was extracted from 6-week-old male wild-type C57BL/6 mice (n=2) and 9-week-old female homozygous hGPR65 knock-in mice (n=2), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers. 

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

Fig.2 Detection of GPR65 expression in hGPR65 knock-in mice by WB. GPR65 was detectable in liver and kidney from both WT C57BL/6 and HO hGPR65 knock-in mice using a cross-actived antibody recognizing human and mouse GPR65 and specific bands were detected for GPR65 at approximately 37 kDa (as indicated). The liver and kidney tissue lysates were collected from 6-week-old male wild-type C57BL/6 mice and 9-week-old female homozygous hGPR65 knock-in mice, and then analyzed by western blot with anti-GPR65 antibody. 

Abbr. HO, homozygous; WT, wild type.

Fig.3 Detection of GPR65 expression in hGPR65 knock-in mice by WB. GPR65 was detectable in kidney and lung tissues from both WT C57BL/6 and HO hGPR65 mice using a cross-actived antibody recognizing the extracellular fragment of human and mouse GPR65, and specific bands were detected for GPR65 between 70–100 kDa, which might be caused by different post-translational modifications. The kidney, lung and spleen tissue lysates were collected from 8-week-old male wild-type C57BL/6 mice and 8-week-old male homozygous hGPR65 knock-in mice, and then analyzed by western blot with anti-GPR65 antibody. 

Abbr. HO, homozygous; WT, wild type.