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. 2021 Dec 3;22(23):13084. doi: 10.3390/ijms222313084

Figure 8.

Figure 8

Involvement of AP-1 and NF-κB in GPR55-mediated Mac-1 transcription. (A,B) THP-1 cells were transfected with luciferase reporter constructs containing AP-1 or NF-κB consensus binding sites, pretreated with the indicated concentrations of CID16020046 for 1 h, and then treated with 10 μM O-1602 for 24 h. The AP-1 (A) and NF-κB (B) luciferase activities were represented as the mean ± SEM from five to six independent experiments. (C,D) Binding of AP-1 and NF-κB to the Mac-1 promoter was detected with the chromatin immunoprecipitation (ChIP) assay. Immunocomplexes of AP-1 and NF-κB associated with DNA were obtained from THP-1 cells. The THP-1 cells were pretreated with the indicated concentrations of CID16020046 for 1 h, and then treated with 10 μM O-1602 for 24 h. Specific DNA fragments were quantified by PCR, as detailed in the Methods section. DNA purified from lysates incubated without antibody was used as input control (Input). Representative images (C) and quantitated histograms of the ChIP assay. The results are presented as means ± SEM of three independent experiments. Statistical significance: *** p < 0.001 vs. non-treated control cells, ## p < 0.01 and ### p < 0.001 vs. O-1602-treated cells.