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. 2020 Jan 17;10:3003. doi: 10.3389/fimmu.2019.03003

Figure 5.

Figure 5

CiTLR22a and CiTLR22b show the opposite effect on IFN and NF-κB pathways by Dual-Luciferase Reporter Assay System and qRT-PCR. (A–G) CIK cells were seeded in 24-well plates overnight and co-transfected with 380 ng of pCiTLR22a-myc, pCiTLR22b-myc, or empty vector; 380 ng of each target plasmid (pIRF3pro-Luc, pIRF7pro-Luc, pIFN1pro-Luc, pIFN3pro-Luc, pIFNγ2pro-Luc, pNF-κB1pro-Luc, or pNF-κB2pro-Luc); and 38 ng of the pRL-TK control reporter vector. Twenty-four hours later, the cells were stimulated with poly(I:C) or unstimulated. The luciferase activities were examined at 24 h post-challenge. (H) CIK cells were transfected with pCiTLR22a-myc, pCiTLR22b-myc, or empty vector in 12-well plates. Twenty-four hours later, the cells were challenged with poly(I:C) for 24 h. Then the cells were harvested for qRT-PCR to quantify the relative expression levels of IRAK3. All the experiments were repeated in triplicate. *p < 0.05, **p < 0.01, ***p < 0.001.