Figure 4

LXR activation is associated with histone deacetylation and inhibition of p50 binding at the Nos2 promoter. A AcH4 ChIP at the Nos2 promoter for LPS-stimulated BV2 cells pretreated with 5 μM GW3965 (GW/LPS) compared to unstimulated (VC) and vehicle pretreated (VC/LPS) cells. Data shown for primers amplifying −551 to −359 upstream of Nos2. B NF-kappaB1 p50 ChIP at the Nos2 promoter for LPS stimulated BV2 cells pretreated with 5 μM GW3965 (GW/LPS) compared to unstimulated (VC) and vehicle pretreated (VC/LPS) cells. Data shown for primers amplifying −159 to −15 bases upstream of Nos2. C Microglial cell line BV2 were pretreated with 1 μM GW3965 (GW), HDAC inhibitor trichostatin A (TSA, 10 nM) or vehicle (VC, 0.1% DMSO) for 1 h, then stimulated with LPS (10 ng/ml). Nitric oxide (nitrite) production was measured by Griess assay. Data are mean + SD. N = 4 independent experiments. D Primary murine microglia were transfected with HDAC3 siRNA (HDAC3 siR) or non-targeting siRNA (NT siR); 72 h following transfection, cells were pretreated with 1 μM GW3965 (GW) or vehicle (VC, 0.05% DMSO) for 1 h followed by stimulation with LPS (10 ng/ml). Nos2 gene expression (relative expression) was determined by real-time RT-PCR. Data are mean + SD. N = 3 independent experiments.