Strain specific analysis of GPR84 mRNA expression in wild-type C57BL/6N mice, B-hGPR84 mice and B-GPR84 KO mice by RT-PCR. Brain RNA were isolated from wild-type C57BL/6N mice (+/+), homozygous B-hGPR84 mice (H/H) and B-GPR84 KO mice (Δ/Δ), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human GPR84 primers. Mouse Gpr84 mRNA was detectable only in wild-type C57BL/6N mice. Human GPR84 mRNA was detectable only in homozygous B-hGPR84 mice. 

Western blot analysis of GPR84 protein expression in homozygous B-hGPR84 mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hGPR84 mice (H/H), and then analyzed by western blot with anti-GPR84 antibody. (anti-human GPR84 antibody: Santa Cruz, sc-293447). 40 μg total proteins were loaded for western blotting analysis. GPR84 was detected in lung, liver, heart and bone marrow, as the antibody is cross-recognized between human and mouse GPR84. (There was no β-actin in the heart tissue, there was no GAPDH in the bone marrow tissue.)