Extended Data Fig. 5: OAA positively regulates antiviral immune activation via ETS2.
From: Oxaloacetate sensing promotes innate immune antiviral defence against influenza virus infection
a, Monocytes/macrophages and DCs identified by scRNA-seq from data deposited to the Gene Expression Omnibus (GEO) under accession number GSE243629. The UMAP projection from healthy controls and infected adults. b, Dot plots displaying marker gene expressions for pDCs, cDC2, CD14+ monocytes and CD16+ monocytes. c, The knockdown efficiency of the siRNAs in THP-1-derived macrophages was detected by qPCR. d, ELISA of IFNβ in PBMCs transfected with Scramble (Scr) or ETS2-specific siRNAs infected with H1N1 (PR8) virus (MOI = 1) for 24 h in the presence of diethyl-oxaloacetate (DEOAA) (1 mM). e, A549 cells transfected with Scr or ETS2-specific siRNAs were challenged with H1N1 (PR8) virus (MOI = 1) in the absence or presence of DEOAA (1 mM). The transcription levels of IFNB1 and ISGs were analyzed by qPCR. f, Intracellular NP vRNA levels in the sample as (e). g, The virus titers of A549 cells transfected with Scr or ETS2-specific siRNAs were infected with H1N1 (PR8) virus (MOI = 1) for 24 h in the presence of diethyl-oxaloacetate (DEOAA) (1 mM). h, 293 T cells transfected with Scr or ETS2-specific siRNAs, together with plasmid of TBK1-Luc. The cells were treated with DEOAA (1 mM) and the protein extracts were harvested for luciferase assay. i, Upper panel: violin plots comparing the antiviral response scores in different cell subpopulations between IAV-infected individuals and healthy controls. Single-cell antiviral response scores were determined using the AddModuleScore function in the R toolkit of Seurat to measure the average expression of a predefined set of antiviral genes in each cell. Lower panel: bar plots comparing the ETS2 regulon activities within myeloid subpopulations between IAV-infected individuals and healthy controls. The ETS2 regulon values were deduced using the runSCENIC_3_scoreCells function from the SCENIC toolkit to calculate the enrichment of the putative target gene set of ETS2 as an area under the recovery curve across the ranking of all genes in a particular cell, whereby genes were ranked by their expression values. c–h, Data are represented as mean ± s.e.m. (n = 3 independent biological experiments). P values were determined using two-tailed unpaired Student’s t test (c, d and f–i) or two-way ANOVA with Tukey’s multiple-comparison test (e).
