Fig. 4.

Transgenic overexpression of ABHD5 did not alter the systemic response to bacterial endotoxin. WT and ABHD5 Tg mice were maintained on a standard chow diet and received a single intraperitoneal injection of either saline or LPS (5 μg per mouse) 1 h prior to dissection of tissues. (A, B) Q-PCR analyses of epididymal white adipose tissue (WAT) (A) and hepatic (B) gene expression. TNFα, tumor necrosis factor α; IL-6, interleukin 6; IL-1β, interleukin 1β; IFNγ, interferon γ. Data in panels (A) and (B) represent the mean ± SD (n = 4); values not sharing a common superscript differ significantly (P < 0.05). (C) Freshly isolated thioglycolate-elicited macrophages were pooled from n = 5 mice per genotype and cultured as described in “Materials and Methods.” To examine TLR4-driven signal transduction, macrophages were treated with 10 ng/ml Kdo₂-Lipid A (TLR4 agonist) for 2 h. Cell samples were collected at 15 and 30 min (15′, 30′) and 2 h (2h), and signaling was examined by immunoblotting to detect phospho-IκBα (Ser32), phospho-SAPK/JNK (Thr183/Tyr185), phospho-p38-MAPK (Thr180/Tyr182), and β-actin as a loading control.