α-SYN-induced NF-κB activation in WT mouse primary microglia. (A) 24 h and 72 h after the treatment of either vehicle or 500 nM aggregated human α-SYN, cells were stained for NF-κB p65 protein (Red) and SYTOX Green to show the nucleus. At both time points, α-SYN-treated microglia exhibited increased immunoreactivity for NF-κB p65, and the nuclear accumulation of NF-κB p65 was quite distinct compared with the vehicle treated ones. Scale bar=40 μm. Arrows indicate the enrichment of nuclear NF-κB p65. (B, C) Quantification of 24 h and 72 h nuclear NF-κB p65 intensity. At least four images from each group were analyzed using ImageJ software. At both time points α-SYN-treated WT microglia had markedly enhanced nuclear NF-κB p65 staining compared with the vehicle-treated controls. *P<0.05, α-SYN vs. vehicle, t-test.