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A–C
293T cells were transfected with an ISRE or IFN‐β promoter reporter plasmid and pRL‐TK plasmid, together with an empty vector (EV) or NLRP11 construct for 24 h, and then transfected with poly(I:C) (5 μg/ml) (A), poly(dA:dT) (5 μg/ml) (B), or infected with Sendai virus (SeV) (MOI = 0.1) for 20 h (C), followed by ISRE‐ or IFN‐β‐dependent luciferase activity (fold induction) analysis. The data were normalized by using the values of ISRE‐luc or IFN‐β‐luc divided by the values of TK‐luc, and then, the results of each group were analyzed to compare with the control group.
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D
293T cells were transfected with the IFN‐β promoter reporter plasmid and pRL‐TK plasmid, together with an empty vector or cGAS and STING plasmids and increasing amount of NLRP11 for 24 h, and analyzed for IFN‐β‐dependent luciferase activity (fold induction).
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E
Immunoblot analysis of the total and phosphorylated (p‐) IRF3 in THP‐1 cells stably transduced with recombinant lentivirus expressing empty vector or shNLRP11‐#1, which were left untreated or infected with SeV (MOI = 1) for indicated time points. Numbers between two blots indicate densitometry of phosphorylated proteins relative to that of total proteins, respectively.
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F, G
Expression of IFNB1, ISG54, and ISG56 mRNA in NLRP11 overexpressing THP‐1 cells (F) or NLRP11‐knockdown THP‐1 cells (G) infected with SeV (MOI = 1) for indicated time points.
Data information: Data in (A–D, F, and G) are expressed as means ± SEM of three independent experiments (*
P < 0.05, **
P < 0.01, and ***
P < 0.001, versus cells transfected with EV with the same treatment, Student's
t‐test, ns: no significant).
Source data are available online for this figure.
