Fig. 5.

The 17β-estradiol enhances the expression of S100-A9. Quantitative real-time PCR analysis showed a higher mRNA expression of S100a9 inK14E7+E₂ and FvB +E₂ mice compared to untreated mice (FvB or K14E7) (A). Bars (relative expression) represent the mean±SD of three independent experiments (*p<0.05, **p<0.01, Student's t-test) normalized to Hprt mRNA and compared with the control (untreated FvB mice). Western blot analysis (B) and immunohistochemistry (C) also revealed a marked up-regulated S100-A9 expression in K14E7+E₂ and FvB +E₂ mice. S100-A9 positive cells in immunohistochemistry show brown staining in the nucleus and cytoplasm. The results show a representative of three experiments, nuclei were counterstained with hematoxylin and visual field at 40 × magnification. Western blot analysis of Human primary and HPV16 E7 transformed keratinocytes treated with vehicle or ICI 182,780 (50 nM for 48 h) (D) and vehicle or E₂ (10 nM) (E) for 24 h after 48 h starvation in the absence of any growth factor. Cell fractionation was conducted, and lysates were subjected to SDS-PAGE followed by Western blot with anti-S100-A9 and anti-beta-actin (control) antibodies. All gene and protein symbols were written according to the HUGO Gene Nomenclature Committee (HGNC) and UniProtKB, respectively.