Figure 7
XL-DNA activates TLR9. A. After HEK293T cells were transfected with NF-κB-luc and pMSCVpuro-mTLR9-Myc-expressing plasmid, cells were treated with XL-DNA. Cell lysates were assayed for luciferase activities. Values are mean ± SEM (n = 3). *, p < 0.05. Representative data from three independent experiments are presented. B. BMDCs isolated from wild-type (WT) or TLR9-knockout (KO) mice were treated with XL-DNA (1 μM), R848 (100 ng/ml), or ODN1668 (ODN; 1 μM). Cell lysates were analyzed for immunoblotting. C, D. BMDCs from WT or TLR9KO mice were treated with XL-DNA (1 μM), R848 (100 ng/ml), or ODN1668 (ODN; 1 μM) for (C) 4 h and (D) 18 h. For C, mRNA levels were determined by quantitative real time-PCR. For D, concentrations of cytokines were measured by ELISA. For A, C, and D, values are mean ± SEM (n = 3). *, p < 0.05. E. Whole cell lysates were obtained from HEK293T cells transfected with mTLR9-Myc/His-expressing plasmid or empty vector and were incubated with biotinylated XL-DNA for 4 h followed by incubation with NA-beads for 2 h. Co-precipitated proteins were analyzed by immunoblotting with anti-Myc antibody. For B-E, representative data from two independent experiments are presented. Veh, vehicle.