Fig. 3

α-Synuclein could induce the microglial secretion of CXCL12 via TLR4/IκB-α/NF-κB signaling. a Real-time qPCR was used to analyze the mRNA expression levels of TLR1, TLR2, and TLR4. b BV-2 cells were preincubated with TAK242 or C29 for 2 h, and the CXCL12 levels in supernatants (500 μl) collected after 12, 24, 36, or 48 h of stimulation with α-synuclein were evaluated by ELISA. c The phosphorylation of IκB-α in BV-2 cells was detected by western blot after 24 h of incubation with α-synuclein with or without TAK242. d, e The levels of NF-κB p65 complex in the nucleus (NF-κB p65 N) and cytoplasm (NF-κB p65 C) were detected by western blot after 24 h of stimulation with α-synuclein with or without TAK242. Actin served as the control for NF-κB p65 C, while histone 3 served as the control for NF-κB p65 N. f BV-2 cells were preincubated with PDTC for 2 h, and the CXCL12 levels in supernatants collected after 12, 24, 36, and 48 h of stimulation with α-synuclein were evaluated by ELISA. The data are shown as the mean ± SEM from three independent experiments. ****p < 0.0001. ***p < 0.001. **p < 0.01. *p < 0.05. BSA served as a control