Figure 6.

Acute liver failure in FLIPf/f-creERT2 mice is induced through activation of innate immune receptors. (a) Endotoxin levels were determined in the plasma of wt and FLIP^f/f-creERT2 mice at 48 h following injection of tamoxifen. (b) Hepatic mRNAs isolated from wt and FLIP^f/f-creERT2 mice at 48 h post tamoxifen were measured by qRT-PCR for relative TLR4 and MyD88 mRNA expression. (c) After 4 weeks of gut decontamination endotoxin levels in the serum, hepatic mRNA expression of TLR4 and (d) ALT levels was determined at 48 h post tamoxifen. (e) Cell-free DNA was measured photometrically in the plasma of wt and FLIP^f/f-creERT2 mice at 48 h following injection of tamoxifen. (f) Relative mRNA expression of TLR9, STING and TBK1 was determined in liver from induced mice±BMC^wt. (g) Human PMA-differentiated THP1 cells with or without chloroquine pretreatment (10 μM) were stimulated with serum or plasma of induced wt and FLIP^f/f-creERT2 mice. The TLR9 ligand ODN 1826 and plasmid DNA as STING ligand were used as controls. After 18–24 h, the secretion of IL-6 and TNF was measured in the supernatants. Data represent mean±S.E.M. (a–f: n≥6 mice/group, g: of three independent experiments each in triplicate), P-values for wt versus FLIP^f/f-creERT2: *P<0.05, **P<0.01, ***P<0.001 and for FLIP^f/f-creERT2 cells±BMC^wt: ^$P<0.05, ^$$P<0.01, ^$$$P<0.001