Generation of CAGp-ephrin-B2 Tg mice.

(A) The structure of the CAGp-ephrin-B2-pA transgene cassettes used to generate transgenic mice. CAGp indicates the CAG promoter and pA the polyadenylation signal. (B) To detect the transgene, PCR analysis of genomic DNA was performed. Bands of the predicted size were detected in mouse lines 12, 21, and 47 and in the 2 dead founders, no. 51 and no. 52, indicating that they are transgenic. As a positive control, cDNA from E9.5 mouse embryos (RT) was used, while genomic DNA from wild-type mice (W) was used as a negative control. (C) Total RNA was prepared from the ear of F1 offspring of the lines 12, 21, and 47 of CAGp-ephrin-B2 Tg mice, respectively. Twenty micrograms RNA was loaded in each lane, and the blot was probed with mouse ephrin-B2 cDNA. Lines 12 and 21 represent high-expressing lines (arrowhead in C), while line 47 represents a nonexpressing line. There is no transgene expression in the ear tissue of nontransgenic littermates of these lines. An arrow indicates the endogenous ephrin-B2 transcript.