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. 2017 Dec 8;293(5):1745–1755. doi: 10.1074/jbc.RA117.000541

Figure 1.

Figure 1.

cFLIPL inhibits IRF7-induced gene expression independent of IRF3 and IRF5. A and B, 293T cells were co-transfected with 450 ng of pil12p40-luc (A) or 450 ng of pifna6-luc and 50 ng of pRL-TK, 1000 ng of pCI, pIRF3CA, or pIRF7 or 500 ng each of pIRF5 and pTRAF6 (B). Cells were also co-transfected with either 1000 ng of pCI, pcFLIPL, or pVpx (A) or pAIP (B). Cells were incubated for 24 h post-transfection. C, 293T cells were co-transfected with 450 ng of pifna6-luc and 50 ng of pRL-TK, 1000 ng of pCI, or 500 ng each of pIRF7 and pMyD88. Cells were also co-transfected with either 1000 ng of pCI, pcFLIPL, or pIRF7DN. Cells were incubated for 24 h post-transfection. D, 293T cells were co-transfected with 450 ng of pifna6-luc and 50 ng of pRL-TK, 1000 ng of pCI, or 250 ng each of pIRF7, pMyD88, pIKKα, and pTRAF6. Cells were also co-transfected with either 1000 ng of pCI, pcFLIPL, or pIRF7DN. Cells were incubated for 24 h post-transfection. E, HeLa cells were co-transfected with 450 ng of pifna6-luc, 50 ng of pRL-TK, 250 ng of pIFNα, and 1000 ng of pCI, pcFLIPL, or pAIP. 24 h post-transfection, cells were incubated in medium lacking or containing 3 μm CpG-A for 3 h. For all experiments, cellular lysates were examined for luciferase activities. Results are shown as -fold induction of luciferase activity relative to pCI-transfected cells. A portion of each lysate was additionally examined for protein expression by using IB to detect FLAG-tagged cFLIPL, myc-tagged Vpx, or myc-tagged AIP. Luciferase assays are representative of three technical replicates, and all luciferase assays were performed at least three times. Data are expressed as the mean ± S.D. Statistically significant differences in experimental samples versus unstimulated, pCI-transfected cells are denoted (*, p < 0.05).