Fig. 8

Nrf-2/HO-1 pathway is involved in the protection mechanism of Fetuin-A in glutamate-treated microglial cells. A–D Western blot analysis of Nrf-2 expression in cytosolic or nuclear. GAPDH and Histone H3 were used as the loading control. And bar graphs show the results of analysis (by band density analysis) of these proteins (n = 3). E The intracellular localization of Nrf-2 was visualized by confocal microscopy. Scale bar is 20 or 50 μm (n = 5). F Oxidative stress of microglial cells were analyzed by detecting the levels of MDA, GSSG, and GSH (n = 3). G, H Mitochondrial superoxide was detected by immunofluorescence using MitoSox Red staining. Scale bar is 50 μm. The relative immunofluorescence intensity of Fetuin-A was detected with Image-J software (n = 3). I, J Detection of RIPK3, MLKL, p-RIPK3 and p-MLKL was done by immunoblot analysis (n = 3). Data are presented as the means ± SD; *P < 0.05 vs. control, **P < 0.01 vs. control, ^#P < 0.05 vs. Glu group, ^&P < 0.05 vs Glu + Fetuin-A group, and n.s.: no significant difference