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. 2013 Apr 2;8(4):e60277. doi: 10.1371/journal.pone.0060277

Figure 1. characteristics of the PIM1 conditional transgenic mice.

Figure 1

A) Schematic representation of the transgene carrying Pim1. The arrow indicates the generation of tissue specific transgenic mice expressing PIM1 by crossing with mice expressing Cre-recombinase under a tissue specific promoter; the Lox/Stop/Lox cassette is excised allowing transcription. B) Expression of PIM1 in Pim1/PSA-Cre mice. RNA was extracted from different tissues of 10-week-old mice and the specific transgenic Pim1 transcription analyzed by RT-PCR. C) Levels of human and mouse PIM1 mRNAs calculated by quantitative RT-PCR. Graph shows average (± SD) levels of expression in the prostate of human (black) or mouse (grey) PIM1 mRNA of at least two mice per genotype performed in triplicate. Data were normalized to 1 (log 10 = 0) using the levels of mouse PIM1 levels in wild type animals. ND: Not detected. Human PIM1 was not detected in WT nor PTEN-Het mice. D) Level of PIM1 protein in the mice of different genotypes measured by Western blot. The prostate of 2 animals (1 and 2) per cohort was processed to extract total proteins and run in a PAGE to allow further identification by western blot. Transgenic PIM1 was identified by myc-tag present in the transgene. Total PIM1 content was identified with a PIM1 antibody cross-reacting with human and mouse species. Activity was measured as Bad phosphorylation levels at S112. E) Representative pictures of high grade mPIN lesions developed after hormone treatment. To determine the development of mPIN lesions due to hormone treatment, 8-week-old untreated and hormone-treated (for 1 or 2 rounds) mice of each genotype were sacrificed and prostate tissue was obtained (see text for details). H&E staining of prostate tissue was used for mPIN grading. Pictures represent: No treatment: wt) pl-grade 0; tgPim1) pl-grade 1; PTEN-Het) pl-grade 5; tgPim1/PTEN-Het) pl-grade 8; 1 round of hormone treatment: wt) pl-grade 2; tgPim1) pl-grade 4; PTEN-Het) pl-grade 10; tgPim1/PTEN-Het) pl-grade 11; 2 rounds of hormone treatment: wt) pl-grade 6; tgPim1) p-grade 13.