Western blot analysis of ADAM9 protein expression in homozygous B-hADAM9 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM9 mice (H/H), and then analyzed by western blot with species-specific anti-ADAM9 antibody. 40 μg total proteins were loaded for western blotting analysis. ADAM9 protein was detected in the heart, liver, spleen, lung, kidney, brain and thymus of wild-type C57BL/6 mice and homozygous B-hADAM9 mice, as the antibody is cross-recognize both human and mouse ADAM9.
mRNA expression analysis
Strain specific analysis of ADAM9 mRNA expression in wild-type C57BL/6 mice and B-hADAM9 mice by RT-PCR. Brain and lung RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM9 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human ADAM9 primers. Mouse Adam9 mRNA was detectable only in wild-type C57BL/6 mice. Human ADAM9 mRNA was detectable only in homozygous B-hADAM9 mice but not in wild-type mice. And the PCR products were confirmed with sanger sequencing.