Fig. 1. Transgene integration, expression and activity. A, Southern analysis of wild-type and transgenic mice. Genomic DNA was isolated from the tails of weanlings, digested with BamHI, fractionated on a 0.7% agarose/Tris-acetate/ethidium bromide gel, and hybridized with a radiolabeled 8-LOX cDNA probe. Lane 1 contains digested genomic DNA from wild-type mice and Lanes 2–7 contain digested genomic DNA from loricrin 8-LOX transgenic mice. The loricrin 8-LOX transgene has a length of 4.7 kb. B, Northern analysis of skin from wild-type and transgenic mice. Total RNA was extracted from the dorsal skin of wild-type and transgenic mice as described in "Materials and Methods." Ten µg of RNA was electrophoresed on a 1% agarose formaldehyde gel, blotted overnight onto a nylon membrane, and hybridized with a radiolabeled 8-LOX cDNA probe; stripped blots were probed with cDNA for GAPDH, as a control for gel loading. C, immunostaining of protein with 8-LOX antibody. Total protein was isolated from wild-type and loricrin 8-LOX transgenic skin as described under "Materials and Methods." Fifteen µg of total protein was fractionated on a 10% SDS polyacrylamide gel. The protein was transferred overnight to a polyvinylidene difluoride membrane and immunostained with the 8-LOX antibody as described previously (2) . Wild-type mice: M590, M591, M299; transgenic mice M351, M528, M179. Positive [Lox Std, 15-Lox-2 (protein)] and negative [12R Lox (protein)] controls are included for verification of antibody specificity (2) .