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. Author manuscript; available in PMC: 2011 Feb 21.
Published in final edited form as: Oncogene. 2010 May 31;29(30):4261–4275. doi: 10.1038/onc.2010.197

Figure 1. Generation and characterization of 15-LOX2 transgenic mice.

Figure 1

(A) Schematic diagram of transgene constructs, which contain the ARR2PB promoter, rabbit β-globin second intron sequence, 15-LOX2 or 15-LOX2sv-b cDNA, and β-globin-SV40 hybrid polyA tail. The sizes (bp) of each module and restriction enzyme sites (K, Kpn I; A, Apa I; X, Xba I; S, Sal I; C, Cla I; B, BamH I, E, EcoR I; N, Nhe I) are indicated.

(B) Prostatic lobes were dissected from animals of the indicated genotypes and ages (m, month), lysed in RIPA buffer and used in SDS-PAGE (50 μg lysate/lane). Immunoblotting was performed using the rabbit polyclonal anti-15LOX2, which recognizes most splice variants including 15-LOX2sv-b. The blots were stripped and reprobed for β-actin.

(C) Prostate-specific transgene expression. Urogenital and other organs indicated were isolated from 2.5-month fl26 15-LOX2 transgenic mice and protein lysates (50 μg/lane) used in Western blotting for 15 LOX2 and GAPDH (loading control).

(D-E) Transgene expression in young and old animals. Representative images of whole-mount prostate sections (without AP) made from 2.4 (D) or 16.5 (E) month (m) old wt, fl26 and svb9 mice (n>15/genotype), stained for HE or 15-LOX2. Images were taken with a Nikon stereomicroscope (Bar = 1 mm). The orientation of the whole-mount images was illustrated in D, panel a (U, urethra).

(F) Transgene expression in young (2.5 month) and old (14.2 month) prostate lobes of fl26 mice.

(G) 15(S)-HETE levels in wt and transgenic VPs measured in the presence of 50 μM AA. Bars represent the mean ± S.D of the measurements obtained from 5 animals/group. *p < 0.05 and **p < 0.01.