Production and characterization of ARO-iCre transgenic mouse line.

<p><b>(a)</b> Schematic diagram of the DNA construct used for producing ARO-iCre mice. The mammalian codon-improved Cre recombinase (iCre, 1051 bp; white box) followed by an SV40 poly(A) signal cassette (right grey box) is controlled by 304 bp of the human Cyp19/aromatase(IIa)_-278 promoter (left grey box). Arrows indicate position and direction of primers. Oligonucleotide sequences are in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0119392#pone.0119392.t001" target="_blank">Table 1</a>. <b>(b)</b> Cre excision is restricted to ovaries (Ov) and testes (Te) of iCre^+/0;IGF1R^flox/WT mice. The excision was only observed in gonads, not in other tissues (He, heart; Mu, muscle; Lu, lung; Sp, spleen; Br, brain; Li, liver; Pa, pancreas) from males (M) and females (F). RT-, negative control (testis cDNA without RT). PCR detected wild-type (W, 256 bp) and floxed (F, 312 bp) alleles in all tissues, and the excised allele (E, 204 bp) exclusively in gonads. <b>(c)</b> Cre recombinase activity detected in somatic cells of testes in adult ARO-iCre males. After crossing the ARO-iCre mice with a ROSA26 Cre reporter mouse, β-galactosidase activity was detected in both SC and LC. No activity was present in germ cells. Right micrograph, low magnification; left micrographs, details in higher magnification. ST, seminiferous tubules. In, interstitium. Arrow head points to SC. Bar represents 50 μm.