Fig. 1

Experimental design. a Rats were fed an ammonia-containing diet for 4 weeks and sacrificed by perfusion for in vivo immunohistochemical analysis of microglial and astrocyte activation (blue box), the protein expression of nuclear NF-kB and TNF-a in glia and Purkinje neurons (green box), and the mRNA expression of TNF-a in glia and Purkinje neurons (violet box) in the cerebellum. The effects of peripheral intravenous infliximab administration were assessed by injecting the drug into the rats once per week (5 mg/kg) starting 3 days before the administration of an ammonia-containing diet. b Rats were fed an ammonia-containing diet and sacrificed at different time points for in vivo analysis of microglial and astrocyte activation after 1 week of hyperammonemia (blue box), in vivo analysis of nuclear NF-kB and TNF-a protein expression in glia and Purkinje neurons after 1 and 2 weeks of hyperammonemia (green box), and ex vivo analysis of nuclear NF-kB and TNF-a protein expression in glia and Purkinje neurons after 2 and 4 weeks of hyperammonemia (red box). c TNF-a expression in Purkinje neurons was also analyzed by immunohistochemistry on postmortem cerebellar samples from patients who died with liver cirrhosis