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. 2022 Jul 8;8(27):eabq0084. doi: 10.1126/sciadv.abq0084

Fig. 4. LYN controls tyrosine phosphorylation and activity of IRFs.

Fig. 4.

(A) Luciferase reporter assay in HEK293T cells transfected with ISRE (IFN-stimulated response element) reporter vector, IRF5-WT, IRF5-S/D active mutant (S434,436,439,449D), SPOP, and constitutively active IKKβ (IKKβ-EE). (B) IP/IB analysis based on control (Ctrl-sg) and SPOP-deficient (Spop-sg) RAW264.7 cells stimulated with CpG-DNA for 1 hour. (C) IP/IB analysis based on control (Ctrl-sg) and SPOP-deficient (Spop-sg) RAW264.7 cells stimulated with CpG-DNA for 1 hour. (D) IP/IB analysis based on RAW264.7 cells with CRISPR-Cas9–mediated deletion of indicated genes. (E and F) Luciferase reporter assay in HEK293T cells transfected with ISRE reporter vector (E) or NF-κB reporter vector (F) plus IRF and LYN expression vectors (0.01 and 0.03 μg). Onefold luciferase activity corresponds to reporter activity without IRF. (G) IP/IB analysis of CpG-DNA–stimulated macrophages derived from MyD88−/− multipotent progenitor cells that were reconstituted with wt or mutant MyD88 [MyD88-AA (SS136/137AA)]. Data represent mean ± SD from three independent experiments. *P < 0.05, ***P < 0.005 are determined by two-way ANOVA with Sidak’s multiple comparison test (A, E, and F).