Gene targeting strategy for B-hAOC3 mice. The exons 1~4 of mouse Aoc3 gene that encode the extracellular domain were replaced by human AOC3 exons 1~4 in B-hAOC3 mice.

Strain specific analysis of AOC3 mRNA expression in wild-type C57BL/6N mice and B-hAOC3 mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6N mice (+/+) and homozygous B-hAOC3 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human AOC3 primers. Mouse Aoc3 mRNA was detectable only in wild-type C57BL/6N mice. Human AOC3 mRNA was detectable only in homozygous B-hAOC3 mice but not in wild-type mice.

Western blot analysis of AOC3 protein expression in homozygous B-hAOC3 mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hAOC3 mice (H/H), and then analyzed by western blot with anti-AOC3 antibody. 40μg total proteins were loaded for western blotting analysis. AOC3 was detected in heart, spleen, kidney, lung and stomach of wild-type C57BL/6N mice and spleen, lung and stomach in homozygous B-hAOC3 mice.