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The paraffin-embedded skin specimens were sectioned at 5?��m, deparaffinized and stained with Masson-trichrome for the visualization of collagen fibres and also processed with haematoxylin and eosin staining for the light microscopic Sunitinib purchase evaluation. The stained tissue sections were examined using an optical microscope AXIOIMAGER (Zeiss, Germany) and five images (200��) were taken per section. Epidermal thickness was determined as the distance from the basal layer to the stratum granulosum/stratum corneum junction. The thickness was measured in each photograph at 10 random sites. The cytokine levels of IL-1�� and IL-6 in the UV-B-irradiated SKH-1 hairless mouse skin tissue were determined by using ELISA. The skin tissue samples were homogenized in tissue protein reagent buffer containing 1%��-mercaproethanol, 1?mol/l ��-glycerophosphate, 0.1?mol/l Na3VO4, 0.5?mol/l NaF with protease inhibitor cocktail and homogenates were centrifuged at 2000?g for 10?min. After the centrifugation, IL-1�� and IL-6 in the tissue supernatants were measured using ELISA kits (R&D Systems, Minneapolis, MN, PI3K Inhibitor Library research buy USA). The protein concentrations of tissue supernatants were determined and the levels of IL-1�� and IL-6 in the skin tissues were normalized and expressed as ��g/g tissue protein. For immunohistochemical analyses, paraffin-embedded integument sections (5-��m thick) were employed. The sections were placed on glass slides, deparaffinated and hydrated Fossariinae with xylene and graded alcohol. The tissue sections were then subjected to incubation with 0.5% H2O2 in methanol for the removal of endogenous peroxidase. The non-specific antibody binding site was blocked by using 3% BSA in PBS-T for 1?h, followed by 1?h incubation with goat anti-mouse SR-A (1:100) or goat anti-mouse ICAM-1 (1:100). The tissue sections were incubated for 1?h with peroxidase-conjugated anti-goat IgG (1:200). The integuments were developed with 3,3��-diaminobenzidine as a substrate for 1?min and counterstained with haematoxylin. The results are presented as mean?��?SEM for each treatment group. Statistical analyses were conducted using Statistical Analysis Systems. Significance was determined by one-way ANOVA followed by Duncan range test for multiple comparisons and were considered significant at P?