Breeding strategy and deletion confirmation for prostatic knockout of Klf5 and Pten. (A) Schematic of breeding design for the generation of mice with all six desired genotypes of Klf5 and Pten using mouse strains of floxed Klf5 (Klf5F/F), floxed Pten (PtenF/F) ,and PB-Cre4. F and W indicate floxed and wild-type alleles, respectively. (B and C) Detection of Klf5 protein expression by IHC staining in 6-month-old APs (B) and Klf5 mRNA expression by real-time RT-PCR in the whole prostate of 6-month-old mice (C) with different Klf5 deletion status and Pten homozygous deletion. (D and E) Detection of Pten protein expression by IHC staining in 6-month-old APs (D) and Pten mRNA expression by real-time RT-PCR in the whole prostate of 6-month-old mice (E) with different Pten deletion status. + and − indicate wild-type and deleted Klf5 or Pten alleles, respectively; * and ** indicate P < .05 and P < .01, respectively, based on Student's t test.