Extended Data Fig. 8: Dynamic OAA production caused by ACLY modulation contributes to antiviral responses.
From: Oxaloacetate sensing promotes innate immune antiviral defence against influenza virus infection
a, Schematic diagram depicting the production of cellular OAA. b, The knockdown efficiency of the siRNAs was detected by qPCR in THP-1 cells transfected with Scr, ACLY-, GOT1-, MDH2- or PC-specific siRNAs. c, The cellular OAA levels in THP-1 cells transfected with Scr, ACLY-, GOT1-, MDH2- or PC-specific siRNAs in the absence or presence of DEOAA (1 mM). d, THP-1 cells transfected with Scr, ACLY-, GOT1-, MDH2- or PC-specific siRNAs were infected with H1N1 (PR8) virus (MOI = 1) for 24 h in the absence or presence of DEOAA (1 mM). The total RNA was harvested and the transcription levels of IFNB1 were analyzed by qPCR. e, Intracellular NP vRNA levels in the sample as (d). f, Schematic diagram of Acly knockout strategy. Deletion of exon 3 and exon 6 results in frame shift and disrupts its open reading frame, leading to the loss of Acly expression. g, Activated macrophages enriched from lung tissues of Aclyfl/fl and Aclyfl/flLyz2Cre mice (n = 3 per group) given intranasal H1N1 (PR8) virus inoculation for 3 d, and the cell lysates were analyzed by immunoblotting with indicated antibodies. h, OAA levels in PBMCs cultured with MEDICA16 (200 μM) or BMS-303141 (200 μM) for 12 h with or without DEOAA (1 mM) treatment. i, ELISA of IFNβ in the supernatant of PBMCs infected with H1N1 (PR8) virus (MOI = 1) in the presence of DEOAA (1 mM), along with MEDICA16 (200 μM) or BMS-303141 (200 μM). j,k, OAA content (j) and qPCR analysis of Tbk1 (k) in BMDMs from Aclyfl/fl and Aclyfl/flLyz2Cre mice cultured with DEOAA (1 mM). b–e,h–k, 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.
