FIGURE 6.

Activated NF-κB pathway and production of proinflammatory cytokines and chemokines in the skin of dCKO and FIP200 CKO mice. A, lysates were prepared from the skin of p53 CKO (lanes 1-3) or dCKO (lanes 4-6) mice and analyzed by Western blotting using anti-phospho-NF-κB p65 (upper), anti-NF-κB p65 (middle), or anti-vinculin (lower) antibodies. B, two-color immunofluorescence of skin sections from p53 CKO (left) or dCKO (right) mice using fluorescein isothiocyanate-conjugated anti-CD45 (green) and Texas Red-conjugated anti-phospho-NF-κB antibodies. Note the CD45⁺ phospho-NF-κB⁺ double-positive leukocytes in the skin of dCKO mice. Sections were counterstained with DAPI (blue). Scale bar = 100 μm. C, skin sections from FIP200^F/F (left) and FIP200 CKO (right) mice were immunostained by fluorescein isothiocyanate-conjugated anti-phospho-NF-κB antibody (green). Sections were counterstained with DAPI (blue). Scale bar = 100 μm. D, semi-quantitative reverse transcriptase-PCR analysis of cytokines and chemokines gene expression for mRNA isolated from the dorsal skin of p53 CKO and dCKO mice. cDNA were used directly (1:1) or diluted 10-fold (1:10) before PCR. Data are representative of more than five independent mouse pairs. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.