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. 2015 Aug 4;5:12738. doi: 10.1038/srep12738

Figure 1. USP18 negatively regulates TLR-induced NF-κB activation.

Figure 1

(a) HEK293T cells were transfected with plasmids of 21 DUBs along with MyD88 and a reporter plasmid carrying the NF-κB promoter (NF-κB-luc). The cells were analyzed for NF-κB activity by a reporter gene assay. (b) Domain organization and immunoblot analysis of human and mouse USP18 proteins. (c) 293T cells were transfected with Flag-MyD88 and NF-κB reporter along with different tagged human or mouse USP18 expression plasmids. The cells were analyzed for NF-κB activity by a reporter gene assay. (d) Myc-USP18 and Flag-MyD88 were transfected in 293T cells and IκBα turnover was monitored by western blotting using indicated antibodies. (e) HeLa cells were transfected with EGFP-USP18 expression plasmid for 48 hrs. Cells were then treated with LPS (100 ng/ml) for 30 min, and then subjected to immunofluorescence analysis using a p65-specific polyclonal antibody. DNA was stained by DAPI (blue). UT, untreated. Scale bar: 10 μm. Data in a,c are presented as the means ± SD of three independent experiments. *P < 0.05, **P < 0.01 and ***P < 0.001, versus cells with the same treatment without USP18 expression (Student’s t-test).