Figure 4

FcγR ^−/− attenuated α-SYN-induced NF-κB signaling and downstream expression of pro-inflammatory molecules. (A) MIP-1α ELISA on the conditioned media collected 2, 4, 8, and 16 h from the vehicle and α-SYN-treated WT primary microglia. At 4 h, but not the other time points, the MIP-1α level was significantly increased with α-SYN treatment compared with the vehicle controls. *P<0.05, α-SYN vs. vehicle, t-test. (B) Conditioned media were collected 4 h and 24 h after the treatment of vehicle or aggregated human α-SYN on WT and FcγR^−/− microglia for a 25-plex mouse cytokine/chemokine assay. At 4 h, MIP-1α and MIP-1β were significantly increased with α-SYN treatment in WT microglia but not FcγR^−/− microglia compared with vehicle-treated ones. All chemokine expression levels were normalized to the level of vehicle-treated WT or FcγR^−/− microglia, respectively. ***P <0.001 WT α-SYN vs. WT vehicle. One-way ANOVA with Tukey’s multiple comparison test. (C) Conditioned media collected from primary WT and FcγR^−/− microglia treated overnight with 100 ng/mL LPS. Both WT and FcγR^−/− microglia respond normally to TLR4 stimulation. TNF expression levels were normalized to the level of vehicle-treated WT or FcγR^−/− microglia, respectively. ***P <0.001 WT LPS vs. WT vehicle, FcγR^−/− LPS vs. FcγR^−/− vehicle. One-way ANOVA with Tukey’s multiple comparison test.