Fig. 1. Generation of conditional and full Tead4 knockout mice. (A) Exon 2 of the mouse Tead4 locus (black boxes) was targeted for homologous recombination. Indicated are the positions of the left arm probe (LAP), right-arm probe (RAP), and binding sites for PCR primers used for genotyping (Table 1), and restriction sites used for ES cell screening and for the insertion of loxP sites (open arrows in targeting vector). After homologous recombination, exon 2 is flanked by loxP sites in the conditional Tead4loxP allele (Cond.). Following Cre-mediated recombination, the Tead4 knockout (KO) allele lacks exon 2. Genomic DNA from ES cells (B) and tail snips (C) was subjected to Southern blotting-hybridization with the LAP. (D,E) Genotypes were routinely confirmed by PCR analysis of tail DNA obtained from the progeny of matings between Tead4^+/- mice showing the absence of Tead4^-/- offspring. Primers: P1, P2, P4 in D and TEAD4.20 (20F), TEAD4.27 (27R), TEAD4.28 (28R) in E. (F) PCR-based genotyping, using primers P1, P2, and P3, of F2 animals produced from mating heterozygotes conditional (Tead4+/lox) animals shows recovery of wild-type, heterozygous and homozygous Tead4 conditional offspring. W, wild type; L, loxP; M, mutant.