Adenosine kinase is a target for the prediction and prevention of epileptogenesis in mice
J. Clin. Invest. Tianfu Li, et al. 118:571 doi:10.1172/JCI33737 [Go to this article.]

Figure 2
Generation and characterization of fb-Adk-def mice. (A) The short isoform of mouse Adk cDNA is located between a human ubiquitin promoter (hUbi) and an SV40 polyA sequence. The UbiAdk transgene is flanked by loxP sites. (B) Adk-Tg mice, which were homozygous for the Adk-KO and the TgUbiAdk transgene, were bred with fb-Adk-def mice, which were heterozygous for Cre. From these crosses, Adk-Tg and fb-Adk-def littermates were produced. (C) Representative PCR of selected animals. Tg, Adk-Tg; def, fb-Adk-def. DNA was amplified with a PCR specific for either the WT (640 bp) or the KO allele (840 bp) of the endogenous Adk gene (Adk-PCR), a PCR specific for the Adk transgene (Tg-PCR), giving rise to a 420-bp amplification product, or a PCR specific for the Cre gene (Cre-PCR), giving rise to a 600-bp product. (D) Representative western blot from adult WT, fb-Adk-def, and Adk-Tg mice. Top: ADK-immunoreactive bands from forebrain (fb) or brainstem plus cerebellum (bs-cb). Bottom: β-Actin–immunoreactive bands were used to normalize for equal loading. (E) Quantitative analysis of ADK levels based on 3 western blots performed with samples from n = 2 animals from each genotype. ADK levels were first normalized to equal loading according to the β-actin standard. These values were then normalized to the respective WT samples (set as 100%). Data represent the mean ± SD of 6 samples. *P < 0.05; **P < 0.01, paired comparisons in t test.