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. 2016 Jun 9;7:11848. doi: 10.1038/ncomms11848

Figure 2. Overexpression of SDC4 inhibits RIG-I-mediated antiviral signaling.

Figure 2

(a,c,d) HEK293 cells were co-transfected with the indicated expression plasmids, luciferase reporter constructs driven by the promoter of gene encoding IFN-β (a), ISRE (c) or NF-κB (d) and pRSV/LacZ as an internal control. Twenty-four hours after transfection, the cells were lysed for luciferase assays (upper panel) and immunoblotting assays (lower panels). (b,e,f) HEK293 cells were transfected with luciferase reporter constructs driven by the promoter of gene encoding IFN-β (b), ISRE (e) or NF-κB (f), and pRSV/LacZ as an internal control. Twenty-four hours after transfection, the cells were infected with SeV for 16 h, and then lysed for luciferase assays (upper panel) and immunoblotting assays (lower panel). (g,h) HEK293 cells were transfected with the indicated expression plasmids. Eighteen hours post transfection, the cells were left untreated or infected with SeV for 6 h, and the cell lysates were analysed by quantitative real-time PCR to examine the levels of the Ifnb1 (g) or Rantes (h) transcripts. (i) HEK293 cells were transfected with the indicated expression plasmids. Twenty-four hours after transfection, the cells were left untreated or infected with SeV for 9 h. The lysates were resolved by native gel electrophoresis (upper panel) or SDS–PAGE (lower panels) and analysed with the indicated antibodies. (j) HeLa cells were transfected with the indicated expression plasmids. Twenty-four hours after transfection, the cells were infected with SeV for 9 h, stained with the indicated antibodies and DAPI, and then imaged by confocal microscopy. Bottom panels were the larger magnification. Scale bar, 50 μM. (k) HEK293 cells were co-transfected with the indicated expression plasmids. Twenty-four hours after transfection, the cells were lysed for luciferase assays (upper panel) and immunoblotting assays (lower panels). The data shown in a–h and k are from one representative experiment of at least three independent experiments (mean±s.d. of duplicate experiments in a–f and k or triplicate experiments in g and h). The two-tailed Student's t-test was used to analyse statistical significance. *P<0.05; **P<0.01; ***P<0.001; NS, not significant versus control groups.