CD97/ADGRE5 Inhibits LPS Induced NF-κB Activation through PPAR-γ Upregulation in Macrophages

Figure 2

Regulation of LPS induced NF-κB activation by CD97 in macrophages. (a) p65 protein level was increased by siRNA mediated CD97 ablation, while p65 expression was inhibited by CD97 ectopic expression (A). CD97 siRNA inhibited CD97 protein level while its ectopic expression increased it (B). (b) CD97 ectopic expression reversed LPS (40 ng/mL) induced p65 expression. (c) CD97 ectopic expression reversed LPS (40 ng/mL) induced p65 nuclear translocation. (d) Different treatments including LPS concentrations and CD97 overexpression modulated NF-κB nuclear activities as analyzed by EMSA. Lane 1: Hela nuclear extract group (positive control). Lane 2: blank control group. Lanes 3 and 4: LPS stimulation promoted NF-κB translocation. Lanes 5, 6, and 7: CD97 overexpression inhibited LPS induced NF-κB translocation. Lanes 8 and 9: CD97 overexpression inhibited the basal NF-κB nuclear level. (e) ChIP assays were performed with LPS stimulation (40 ng/mL) or LPS stimulation and pc-CD97 transfection using antibodies against p65. Chromatin collected before immunoprecipitation served as an input control. The corresponding NF-κB responsive elements in the promoter of TNF-α gene were amplified and analyzed in later qRT-PCRs ().

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