Fig. 5

Sustained hyperammonemia increases the nuclear translocation of the p50 and p65 subunits of NF-KB in Purkinje neurons and glial cells, which is prevented by infliximab. Analysis of NF-kB in neurons was performed by immunofluorescence using antibodies against the p50 (a) and p65 (b) subunits of NF-kB (green staining). Nuclei were stained blue with DAPI. The nuclear/cytoplasmic ratios of p50 (c) and p65 (e) and the proportion of cells containing p50 in the nucleoli (d) are quantified. To analyze the nuclear translocation of NF-kB in glial cells, double immunofluorescence was performed for NF-kB p50 (green in f and red in g) and Iba-1, a microglial marker (red in F), or GFAP, an astroglial marker (green in g). The merged images show the colocalization of these proteins (yellow). The number of microglia (H) and astrocytes (i) expressing nuclear p50 was quantified. The values are the mean ± SEM of 3–4 rats per group. Values that were significantly different from those of the control rats are indicated by asterisks, and values that were significantly different from those of the HA VH rats are indicated by a. *p ≤ 0.05, **p ≤ 0.01, ***p < 0.001, a p < 0.05; aa p ≤ 0.01. C VH = vehicle-treated control rats, C INFLIX = control rats treated with infliximab, HA VH = vehicle-treated hyperammonemic rats, HA INFLIX = hyperammonemic rats treated with infliximab