Gene targeting strategy for B-hNTRK1 mice. The exons 1-10 of mouse Ntrk1 gene that encode the extracellular domain were replaced by human NTRK1 exons 1-10 in B-hNTRK1 mice.
mRNA expression analysis
Strain specific analysis of NTRK1 gene expression in wild-type C57BL/6 mice and B-hNTRK1 mice by RT-PCR. Mouse NTRK1 mRNA was detectable only in cerebellum tissue of wild-type mice (+/+). Human NTRK1 mRNA was detectable only in homozygous B-hNTRK1 mice (H/H) but not in wild-type mice.
Protein expression analysis
Strain specific NTRK1 expression analysis in homozygous B-hNTRK1 mice by western blotting. Cerebellum tissue was collected from wild-type C57BL/6 mice (+/+) and homozygous B-hNTRK1 mice (H/H) and analyzed by western blotting with anti-NTRK1 antibody. Mouse NTRK1 was detectable in wild-type mice and homozygous B-hNTRK1 mice with a known cross-reactive anti-NTRK1 antibody. The detected protein size was 140 kDa, represents a mature form of glycoprotein. Human NTRK1 was also detectable in homozygous B-hNTRK1 mice and wild-type mice with another anti-NTRK1 antibody. So this antibody maybe also a cross-reactive antibody. The detected protein size was about 110 kDa, represents a primary form of glycoprotein.