Fig. 2. Generation of Ctgf^-/- mice. (A) Map of the Ctgf locus (wild-type allele), targeting vector and mutant allele produced by homologous recombination. The HindIII fragments expected for the wild-type (8.6 kb) and mutant (7.6 kb) alleles are indicated by double-headed arrows. A, ApaI; B, BamHI; C, ClaI; H, HindIII; N, NotI; P, PstI; R, EcoRI; S, SmaI; X, XbaI. (B) Southern blot analysis of genomic DNA from a heterozygote intercross. (C) Verification that the targeted locus encodes a null allele. RT-PCR analysis of fibroblasts from wild-type and mutant embryos treated with TGFß1 (5 ng/ml) to induce Ctgf expression. No Ctgf transcripts can be detected in mutants, even after TGFß treatment.