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Fig. 4 Endothelial cell? and secondary heart field cell?specific Adamts16 H355Q mutations recapitulate the BAV phenotype. A, Schematic strategy for the generation of the conditional Adamts16 H355Q mutant knockin mouse line by Cre recombination. B, Schematic showing the location of primers used for reverse transcription-polymerase chain reaction (RT-PCR) analysis of the conditional Adamts16 H355Q mutant knockin mouse line; predicted band sizes are indicated. C, RT-PCR analysis process based on the primer pairs designed as described above for identifying Adamts16+/flox-H355Q;Cre+ mice by detecting a specific 392-bp band of the amplicon of the polymerase chain reaction. The numbers 28 and 65 indicate mouse identity documents, both of which were Adamts16+/flox-H355Q;Cre+ mice according to the RT-PCR result compared with that of a WT mouse. Note that the polymerase chain reaction result for identifying the Cre recombinase is not shown. D, Anatomic analysis of aortic valves in Adamts16+/flox-H355Q;Tie2-Cre+, Adamts16+/flox-H355Q;Wnt1-Cre+, Adamts16+/flox-H355Q;Isl1-Cre+, and Adamts16+/flox-H355Q mice at 3 months of age. The black arrow indicates commissures. Scale bars=1 mm. E, The BAV incidences of Adamts16+/flox-H355Q;Tie2-Cre+ (3 of 30), Adamts16+/flox-H355Q;Wnt1-Cre+ (0 of 39), Adamts16+/flox-H355Q;Isl1-Cre+ (2 oof 26), and Adamts16+/flox-H355Q (0 of 53) mice. P values were calculated using the Fisher exact test. ADAMTS indicates a disintegrin and metalloproteinase with thrombospondin motifs; BAV, bicuspid aortic valve; He, heterozygous; M, marker; and WT, wild-type.