Figure 3

Cultured microglial cells (N9) treated with LPS and 100 nM anti-murine CB ₂ siRNA lose their ability to respond to CB ₂ agonist, JWH-015. A, Microglial cells treated with LPS (100 ng/mL) secreted large quantities of TNF-α (n = 3, **p < 0.005). Co-treatment with JWH-015 (5 μM) attenuated LPS-induced TNF-α release. Pre-treatment with anti-CB₂ siRNA abolished JWH-015's ability to reduce LPS-induced TNF-α release (n = 3, ** p < 0.05). Non-targeting anti-GFP siRNA control had no effect. B and C, Western blot using an anti-murine CB₂ antibody demonstrates that 100 nM anti-CB₂ siRNA significantly reduced expression of CB₂ protein by N9 microglial cells after 48 hr (n = 2, ** p < 0.05).