Figure 3. Conditional endothelial deletion of Nck (noncatalytic region of tyrosine kinase) 1 but not Nck2 blunts partial carotid ligation–induced permeability.

A, Schematic of targeting strategy to construct Nck1 fl/fl mice. Nucleotide substitutions were made in exon 2. Embryonic stem cells transfected with the targeting constructs were screened for homologous recombination by polymerase chain reaction (PCR). PCR showing heterozygous Nck1 fl/+ and homozygous Nck1 fl/fl mice with the expected products at 490 bp (targeted alleles) and wild‐type (Nck1+/+) at 377 bp. B, Pulmonary endothelial cells isolated from different experimental groups after tamoxifen injection and 2 weeks’ recovery, Nck1 and Nck2 expression were assessed by immunoblotting n=2 per group. C, Schematic of the study in which 4 groups of mice were subjected to the ligation surgery as indicated animal genotypes and time of surgery. D, Nck1 but not Nck2 knockout mice showed less Evans blue extravasation and (E) fluorescein isothiocyanate (FITC)‐dextran in ng/μL after ligation, n=5 or 6 per group. Data are expressed as mean±SEM and analyzed by 2‐way ANOVA and Bonferroni posttest (*P<0.05, **P<0.01) cKO indicates conditional knockouts; M, marker; Neo, neomycin; SDA, self‐deletion anchor site.