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. 2012 Aug 27;209(9):1689–1702. doi: 10.1084/jem.20101355

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Figure 3. — IL-1α autocrine signaling contributes to oncogenic RAS-mediated activation of NF-κB–regulated genes. (A) Real-time PCR analysis of Cxcl1, Csf2, Mmp9, and Tnf mRNA expression in control keratinocytes or keratinocytes transduced for 3 d with v-ras^Ha and infected with A-CMV (control) or degradation-resistant IκBα (IκBsr) adenovirus to block NF-κB activity for an additional 2 d. *, P < 0.05 between v-ras^Ha control adenovirus and v-ras^Ha IκBsr adenovirus. (B) Real-time PCR analysis of Cxcl1, Csf2, Mmp9, and Tnf mRNA expression in control or v-ras^Ha–transduced WT keratinocytes treated with PBS or IL-1R antagonist (IL-1ra, Anakinra). *, P < 0.05 between v-ras^Ha PBS and v-ras^Ha IL-1ra. (A and B) Data shown are representative of three independent experiments, and bars represent the mean ± SEM of three replicates. (C) Real-time PCR analysis of Cxcl1 and Csf2 mRNA expression in control and v-ras^Ha–transduced keratinocytes treated with control IgG, IL-1α neutralizing antibodies, or IL-1β neutralizing antibodies. Data shown are representative of two independent experiments, and bars represent the mean ± SEM of three replicates. *, P < 0.05 between v-ras^Ha + IgG and v-ras^Ha + anti–IL-1α. (D) Nuclear extracts (top) or total cell extract (bottom) from primary keratinocytes transduced for 3 d with v-ras^Ha in the presence or absence of IL-1ra (Anakinra) were analyzed by Western blotting. The picture is representative of three independent experiments. Molecular mass is indicated in kilodaltons.