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. Author manuscript; available in PMC: 2013 Oct 30.
Published in final edited form as: Nat Med. 2012 Jul 8;18(8):10.1038/nm.2861. doi: 10.1038/nm.2861

Figure 1.

Figure 1

RNA from UVB-irradiated keratinocytes induces the production of inflammatory cytokines. (a) Quantitative PCR (qPCR) measurements of TNF-α and IL-6 mRNA (24 h) in human keratinocytes cultured for 24 h with control NHEK lysates (nonirradiated), lysates from UVB-irradiated (UVR) NHEKs or lysates from UVR NHEKs first treated with RNase. (b) The concentrations of TNF-α and IL-6 protein measured by ELISA from culture supernatants of NHEKs after the additions described in a. (c) The concentrations of TNF-α and IL-6 protein from PBMCs treated with lysates from UVR NHEKs as described in a. (d,e) qPCR measurements of TLR3 mRNA (24 h) (d) and of TNF-α and IL-6 mRNA (e) from NHEKs treated with TLR3 siRNA or control NHEKs treated with vehicle, transfection reagent (DF) or control oligonucleotides (ctrl siRNA) and then stimulated with lysates from UVR NHEKs. (f) Concentrations of TNF-α and IL-6 protein from culture supernatant of NHEKs measured by ELISA after the experiments described in e. (g) The ears of a wild-type C57BL/6 mouse and a Tlr3-/- mouse 24 h after intradermal ear injection of lysates from UVR NHEKs or equal amounts of lysates from nonirradiated NHEKs. (h) Thickness of the ear skin in mice treated as described in g. (i) qPCR measurements of TNF-α and IL-6 mRNA in tissue extracts of ear skin treated as described in g. To determine statistical significance between groups, comparisons were made using two-tailed t tests *P < 0.05, ***P < 0.001. Data are means ± s.e.m. and are representative of at least three independent experiments. n = 4–6 mice per group. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.