Gene targeting strategy for B-hTSPAN2 mice. The exons 2-8 of mouse Tspan2 gene that encode extracellular domain, transmembrane domain and cytoplasmic portion are replaced by human counterparts in B-hTSPAN2 mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric TSPAN2 expression is driven by endogenous mouse Tspan2 promoter, while mouse Tspan2 gene transcription and translation will be disrupted.

Western blot analysis of TSPAN2 protein expression in homozygous B-hTSPAN2 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTSPAN2 mice (H/H), and then analyzed by western blot with cross reactive anti-TSPAN2 antibody (Proteintech, 20463-1-AP). 40 μg total proteins were loaded for western blotting analysis. TSPAN2 was detected in heart, liver, lung, stomach, urinary bladder and ovary of both wild-type mice and homozygous mice.

Strain specific analysis of TSPAN2 mRNA expression in wild-type C57BL/6 mice and B-hTSPAN2 mice by RT-PCR. Lung and stomach RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hTSPAN2 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human TSPAN2 primers. Mouse Tspan2 mRNA was detectable only in wild-type C57BL/6 mice. Human TSPAN2 mRNA was detectable only in homozygous B-hTSPAN2 mice but not in wild-type mice.