Primer sequences.

<p>Primer sequences.</p

Association between the characteristic phenotype of HPV–associated lesions and genotype of HPV E6/E2 DNA of mice.

<p>Association between the characteristic phenotype of HPV–associated lesions and genotype of HPV E6/E2 DNA of mice.</p

Normalized relative expression of miR-155 in ear and chest normal tissue (a).

<p>Normalized expression of miR-155 in different lesions of K14-HPV16 transgenic mice (CIS and hyperplasia) (b).</p

Normalized relative expression of miR-155 in transgenic mice (HPV+) and wild-type mice (HPV-), in chest (a) and ear tissue (b).

<p>Normalized relative expression of miR-155 in transgenic mice (HPV+) and wild-type mice (HPV-), in chest (a) and ear tissue (b).</p

Mice genotyping.

<p>The presence of integrated HPV was assessed by amplification of HPV-E2 (b) and HPV-E6 (c) genes by polymerase chain reaction methodology (PCR) in-house. Samples 1 and 3 are HPV+; sample 2 is HPV-. Mouse-β-globin gene was used as endogenous control (a). M: molecular weight size marker: (a)100 bp, (b,c)50 bp; C-: negative control.</p

Wild-type (-/-) and K14-HPV16 transgenic (+/-) mice.

<p>Transgenic mice show a hunched position, partial thoracic and cephalic alopecia, together with extensive hyperkeratosis and auricular erythema.</p

Overview of genotyping, histological and miR-155 profiling results.

<p>MiR-155 levels are significantly higher in normal chest skin compared with ear skin samples. Targeted expression of HPV-16 oncogenes to basal keratinocytes leads to multistep skin carcinogenesis–transgenic ear skin samples showed CIS while chest samples showed epidermal hyperplasia. Hyperplastic (chest) skin samples showed a significant miR-155 downregulation compared with matched wild-type samples. No differences were observed between wild-type and transgenic ear samples or between transgenic ear and chest samples.</p

Histology of wild-type and transgenic mice.

<p>a) Wild-type (-/-) female FVB/n mouse. Chest skin showing normal histology. H&E, 200x bar = 50μm b) K14-HPV16 transgenic (+/-) female FVB/n mouse. Chest skin showing epidermal hyperplasia and orthokeratotic hyperkeratosis. Note increased number of epidermal strata with conserved orderly squamous differentiation. c) K14-HPV16 transgenic (+/-) female FVB/n mouse. Ear skin showing in situ carcinoma. Note loss of epidermal stratification and progressive differentiation, presence of suprabasal mitotic figures and anisocytosis and abrupt parakeratotic keratinization with hyperkeratosis. The underlying stroma exhibits intense mixed inflammatory cell infiltration and neovascularization.</p