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. 2009 Aug 3;206(8):1727–1737. doi: 10.1084/jem.20082152

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© 2009 Beraza 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 3. — Hepatocyte-specific deletion of NEMO promotes spontaneous activation of liver NK/NKT cells, and administration of NK1.1-depleting mAb attenuates the damaging phenotype in NEMO^Δhepa mice. (A) FACS analysis revealed a lower number of NK1.1⁺CD3⁻ and NK1.1⁺CD3⁺ cells in NEMO^Δhepa mice. The graph represents the percentage of NK1.1⁺ cells related to the percentage of CD45⁺ cells in the liver (B) FACS analysis of NK1.1⁺/annexin V⁺ related to % of CD45⁺ cells revealed strong apoptosis of NK1.1⁺ cells in livers from NEMO^Δhepa mice. (C) RT-PCR showed strong expression of IL-12 (p35), IFN-γ, and CCL5 but lower IL-4 mRNA in livers. Data are presented as times versus NEMO^f/f untreated. (D) mRNA analysis of isolated NK1.1⁺ cells confirmed cell activation and increased cytokine expression. (E) FACS analysis showed stronger TRAIL expression on NK1.1⁺ cells from livers from NEMO^Δhepa mice. mRNA analysis on isolated NK1.1⁺ cells confirmed this. (F and G) FACS analysis proved effective depletion of NK1.1⁺ cells (F) 40 h after NK1.1 mAb administration that reduced IFN-γ and CCL5 mRNA levels (G). (H–J) Serum ALT (H), H&E staining (I), and FACS analysis (J; CD11b⁺ cells) were used as markers of liver damage and inflammation. Bars, 50 µm. All data are representative of three independent experiments. n = 4. *, P < 0.05; **, P < 0.01; ***, P < 0.001 (NEMO^f/f vs. NEMO^Δhepa). §§, P < 0.01 (NEMO^Δhepa vs. Nk1.1mAb/NEMO^Δhepa). Error bars represent SD.