Figure 5. Validation of inducible renal tubule–specific deletion of S1P.

(A) Schematic illustration of the PCR strategy to detect the floxed and recombined alleles by using primers P1 × P2 and P1 × P3, respectively. (B) PCR amplification using P1 × P2 to detect the floxed allele from various organs of S1P^fl/fl-Cre⁺ and S1P^fl/fl-Cre^– mice following doxycycline treatment. WT denotes C57/BL6 mouse. This amplification resulted in a 434-bp product from the floxed allele and a 380-bp product from WT allele. (C) PCR amplification using P1 × P3 to detect the recombined allele from various organs of S1P^fl/fl-Cre⁺ following doxycycline treatment. The recombined allele was detected as an 1800-bp product. (D) Confirmation of renal S1P deletion at protein level. Immunoblotting analysis of renal S1P protein expression in mice with the indicated genotypes following the same doxycycline treatment.