Fig. 7

Inhibition of oxidative stress is a key pathway for Fetuin-A to repress necroptosis. The microglial cells were treated with 100 μm glutamate (Glu) for 24 ho to induce cellular injury. A, F The oxidative stress of microglial cells were analyzed by detecting the levels of MDA, GSSG, and GSH (n = 3). B, C, 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). D Mitochondrial membrane potential was detected by the JC-1 fluorescence ratio. Scale bar is 50 μm (n = 5). E Cell viability was measured by Cell Counting Kit-8 assay (n = 5). I, J Expression of RIPK3, MLKL, p-RIPK3 and p-MLKL in microglial cells under various treatment conditions. GAPDH was used as the loading control. And bar graphs show the results of analysis (by band density analysis) of these proteins (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, and n.s.: no significant difference