Gene targeting strategy for B-hTYRO3 mice. The exons 2~10 of mouse Tyro3 gene that encode the extracellular domain were replaced by human TYRO3 exons 2~10 in B-hTYRO3 mice.

Strain specific analysis of TYRO3 mRNA expression in wild-type C57BL/6 mice and B-hTYRO3 mice by RT-PCR. Brain RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hTYRO3 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human TYRO3 primers. Mouse Tyro3 mRNA was detectable only in wild-type C57BL/6 mice. Human TYRO3 mRNA was detectable only in homozygous B-hTYRO3 mice but not in wild-type mice.

Western blot analysis of TYRO3 protein expression in homozygous B-hTYRO3 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTYRO3 mice (H/H), and then analyzed by proteinsimple-western blot with anti-TYRO3 antibody. 3μg total proteins were loaded for western blotting analysis. TYRO3 protein was detected in the lung, stomach and brain of wild-type C57BL/6 mice and homozygous B-hTYRO3 mice, with high expression in the brain, as evidenced by cross-reactivity of antibody against mouse and human TYRO3 proteins.