Figure 3. NLRP11 associates with MAVS to inhibit IRF3 activation.

1. Luciferase activity in 293T cells transfected with the ISRE promoter reporter plasmid and pRL‐TK plasmid for 24 h, together with expression plasmids encoding RIG‐I‐CARD, MDA5, MAVS, TBK1, and IRF3 (5D), and increasing amount of NLRP11.
2. 293T cells were transfected with HA‐NLRP11 and Flag‐RIG‐I, Flag‐MDA5, Flag‐MAVS, Flag‐TBK1, and Flag‐IRF3 for 24 h, and cell lysates were subjected to immunoprecipitation with anti‐Flag beads, followed by immunoblot analysis with indicated antibodies.
3. NLRP11 and MAVS interaction was analyzed by co‐immunoprecipitation assay in THP‐1 monocytes upon Sendai virus (SeV) (MOI = 1) infection for 20 h.
4. The domain structure of MAVS. Numbers in parentheses indicate amino acid position in the construct.
5. 293T cells were transfected with Myc‐NLRP11 and Flag‐MAVS, Flag‐MAVS‐ΔCARD, and Flag‐MAVS‐ΔTM for 24 h, and cell lysates were subjected to immunoprecipitation with anti‐Flag beads, followed by immunoblot analysis with indicated antibodies.
6. The domain structure of NLRP11. Numbers in parentheses indicate amino acid position in the construct.
7. 293T cells were transfected with HA‐MAVS and Flag‐NLRP11, Flag‐NLRP11‐PYD, Flag‐NLRP11‐NOD, and Flag‐NLRP11‐LRR for 24 h, and cell lysates were subjected to immunoprecipitation with anti‐Flag beads, followed by immunoblot analysis with indicated antibodies.
8. Luciferase activity in 293T cells transfected with the ISRE promoter reporter, pRL‐TK plasmid, and MAVS or empty vector (EV), together with expression plasmids encoding Flag‐NLRP11‐PYD, Flag‐NLRP11‐NOD, or Flag‐NLRP11‐LRR for 24 h.

Data information: Data in (A and H) are expressed as means ± SEM of three independent experiments (*P < 0.05, **P < 0.01, and ***P < 0.001, versus the cells transfected with EV with the same treatment, Student's t‐test). Source data are available online for this figure.