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. 2016 Jun 28;6:28845. doi: 10.1038/srep28845

Figure 4. Setdb1-deficiency increases NF-κB p65 recruitment to the IL6 promoter.

Figure 4

(a) An IL6 promoter luciferase assay in J774.1 macrophages infected with shSetdb1 or shGFP retrovirus. Cells were transiently transfected with a series of truncated or mutated (indicated with “x”) IL6 promoters and treated with LPS (100 ng/ml). (*P < 0.05, n = 3). (b) A ChIP assay of p65 in peritoneal macrophages from KO and WT mice. Cells were treated with lipid A (100 ng/ml) or vehicle for indicated times. GAPDH gene was used as a negative control. (*P < 0.05 vs. WT, ##P < 0.01 vs. 0 h, n = 3). (c) Representative Western blots for p65 in nuclear extracts from shSetdb1- and shGFP-J774.1 macrophages. Cells were treated with LPS (100 ng/ml) or vehicle for indicated times. (d,e) Representative Western blots for phosphorylated JNK, p38, and ERK and IκBα. The whole cell lysates were used. Peritoneal macrophages from KO and WT mice were treated (d) with the indicated concentrations of lipid A or vehicle for 15 minutes or (e) with 100 ng/ml of lipid A or vehicle for indicated times.