Strain specific analysis of ADAM8 mRNA expression in wild-type C57BL/6 mice and B-hADAM8 mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM8 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human ADAM8 primers. Mouse Adam8 mRNA was detectable only in wild-type C57BL/6 mice. Human ADAM8 mRNA was detectable only in homozygous B-hADAM8 mice but not in wild-type mice.
Protein expression analysis
Western blot analysis of ADAM8 protein expression in homozygous B-hADAM8 mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hADAM8 mice (H/H), and then analyzed by western blot with cross reactive anti-ADAM8 antibody. 40 μg total proteins were loaded for western blotting analysis. The red arrow represent the pro-ADAM8, and the blue arrow represent the active form of ADAM8. The pro-ADAM8 was detected in liver, spleen, lung, kidney and brain, while the active ADAM8 was detected in heart and liver of both wild-type mice and homozygous mice.