Fig. 6

Neuron-derived CCL2 stimulated the activation of NF-κB in microglia. As previously described, the medium and lysates (CM) of neurons stimulated with TNF-α were collected and added into microglia cultures. The microglia was pretreated with 5 μM CCL2 receptor inhibitor (INCB or C021) for 1 h before the addition of CM to block CCL2. After incubation for 24 h, NF-κB activation was evaluated through determination of IκBα phosphorylation and NF-κB p65 nuclear translocation using western blotting (a–c). d The cultured microglia was pretreated with 10 μM PDTG (NF-κB inhibitor) for 1 h prior to administration of CM. After cultured for 24 h, the mRNA expression of IL-6 and IL-1β was measured using qRT-PCR. **P < 0.01, *P < 0.05, vs Ctrl; ^## P < 0.01, ^# P < 0.05, vs CM