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. 2009 Sep 28;206(10):2161–2177. doi: 10.1084/jem.20090616

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© 2009 Kovalenko et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jem.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 8. — Caspase-8–deficient keratinocytes display an enhanced response to transfected DNA. (A and B) Effect on gene activation. (A) Effect of caspase-8 knockdown. Relative expression of the IFNβ gene (ratio of the IFN-β transcript in caspase-8 knockdown keratinocytes to the same transcripts in mock-transfected cells) assayed in keratinocytes from one pair of P1 littermates that were transfected in duplicate, 1 d after plating, with either control siRNA (blue) or caspase-8 siRNA (maroon) and, 72 h later, transfected with poly(dA-dT) for the indicated times. Shown are mean expression ratios ± SD for the respective duplicate transfections. (B) Effect of caspase-8 KO. Expression of the indicated gene transcripts at different times after DNA transfection of keratinocytes from Casp-8^F/+K5-Cre mouse and Casp-8^F/−K5-Cre mouse epidermis from two pairs of P1 littermates, 1 d after plating. Cells were transfected either with poly(dA-dT) or with calf thymus DNA, as described in Materials and methods. (C) Effect on Isg15 expression. Western analysis of the expression of Isg15 protein at different times after poly(dA-dT) transfection of keratinocytes from Casp-8^F/+K5-Cre and Casp-8^F/−K5-Cre epidermis from two pairs of P1 littermates, 1 d after plating, as in A. Analysis of the expression of β-actin served as a loading control. The data presented in each of the panels are representative of findings obtained in two independent experiments.