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. 2015 Feb 27;11(4):434–447. doi: 10.7150/ijbs.9311

Figure 1.

Figure 1

Ttc9a transgene construct and screening strategy. (A) Strategy for targeted disruption of Ttc9a gene. Ttc9a exon1 was replaced by neomycin cassette by homologous recombination as indicated by the dotted lines in the mouse ES cells. Exons are indicated by filled boxes; homology arms are indicated by thick lines; introns are indicated by thin lines; and loxp sites are indicated by triangles. (B) Southern blot analysis of Sca I-digested ES cell and tail genomic DNA from wild (+/+), heterozygote (+/-) and homozygous (-/-) Ttc9a KO mice. 5' probe detect a ~11 kb wild-type (Wt) and a ~4 kb mutant (Mut) band. (C) Genotyping tail DNA by PCR screening. The wild-type allele yielded a 200 bp band size and the mutant allele gave 654 bp. (D) TTC9A protein expression is abolished in Ttc9a-/- mice. Total protein extract from mice tissues (brain, mammary gland (Mg), lung and uterus) were analysed by immunoblotting with TTC9A specific antibody. GAPDH was used as a loading control. The blot shows complete absence of TTC9A protein in the 4 tissues assayed from Ttc9a-/- mice.