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. 2018 Jul 18;9:1509. doi: 10.3389/fimmu.2018.01509

Figure 4.

Figure 4

TAARD increases the luciferase reporter activities of NF-κB and STAT3 via TLR1 and TLR3, respectively. (A) NKL cells were electroporated with either pGL3-κB-Luc (1 µg) or pGL3-Basic (1 µg) and a pRL-TK renilla-luciferase control plasmid (50 ng). After electroporation, cells were immediately transferred into fresh medium and cultured for an additional 6 h. Then, cells were treated with different concentrations of TAARD. (B) NKL cells were electroporated as described in (A) but with 4×M67 pTATA TK-Luc plasmid (1 µg). Cells were treated and measured as described in (A). (C) 293T cells were co-transfected with pGL3-κB-luc (1 µg) or pGL3-Basic (1 µg) and pRL-TK renilla-luciferase control plasmids (5 ng) in the presence or absence of a TLR1 (0.5 µg) expression plasmid by Lipofectamine 2000. Twenty-four hours later, cells were treated with different concentrations (0.1, 1, and 10 µM) of TAARD for another 24 h with fresh medium. (D) 293T cells were co-transfected with 4×M67 pTATA TK-Luc (1 µg) or pGL Basic plasmid (1 µg) and pRL-TK renilla-luciferase control plasmids (5 ng) in the presence or absence of a TLR3 (0.5 µg) expression plasmid by Lipofectamine 2000. Twenty-four hours later, cells were treated with different concentrations (0.1, 1, and 10 µM) of TAARD for another 24 h with fresh medium. The ratio of firefly to renilla luciferase activities was used to show the relative luciferase activity, which corresponded to NF-κB or STAT activation. Data shown represent one of three independent experiments with similar results. *p < 0.05, **p < 0.01, and ***p < 0.001 denote statistical comparison between the two marked treatment groups.