Targeting strategy
Gene targeting strategy for B-hTNC mice. The exons 2-27 of mouse Tnc gene that encode the full-length protein were replaced by human TNC exons 2-28 in B-hTNC mice.
Protein expression analysis
Western blot analysis of TNC protein expression in homozygous B-hTNC mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTNC mice (H/H), and then analyzed by western blot with cross-reactive anti-TNC antibody. 30 μg total proteins were loaded for western blotting analysis. Two isoforms of TNC were detected in thymus and lung.
mRNA expression analysis
Strain specific analysis of TNC mRNA expression in wild-type C57BL/6 mice and B-hTNC mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hTNC mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human TNC primers. Mouse Tnc mRNA was detectable only in wild-type C57BL/6 mice. Human TNC mRNA was detectable only in homozygous B-hTNC mice but not in wild-type mice.
