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. 2024 May 2;63(5):2301232. doi: 10.1183/13993003.01232-2023

FIGURE 4.

FIGURE 4

Gasdermin B (GSDMB) binds to mitochondrial antiviral-signalling protein (MAVS). a) Expression of OASL in control or GSDMB knockout (KO) BEAS-2B cells transfected with either empty vector (EV) or Flag-tagged TANK binding kinase 1 (TBK1), followed by high molecular weight (HMW) polyinosinic-polycytidylic acid (poly(I:C)) transfection for 6 h. b) Co-immunoprecipitation (IP) studies: anti-haemagglutinin (HA) immunoblots of either input or immunoprecipitated (IP:Flag) protein complex with anti-Flag from HEK293T cells transfected with HA-GSDMB and either Flag-MAVS, Flag-TBK1 or Flag-interferon regulatory factor 3 (IRF3). c) Schematic illustration of HA-GSDMB constructs used in co-IP assays in d. d) Flag-MAVS co-immunoprecipitated with various domains of HA-GSDMB in HEK 293T cells. e) Expression of OASL in control or GSDMB KO BEAS-2B cells transfected with EV or Flag-MAVS, followed by HMW poly(I:C) transfection for 6 h. f) Expression of OASL in GFP-overexpressing or GSDMB-overexpressing cells with MDA5 depletion, followed by transfection of HMW poly(I:C) for 6 h. For a, e, f, mean±sems shown from at least three independent biological replicates. For all immunoblot data, representative results are shown from two independent biological experiments. NT: no treatment; ns: nonsignificant; sh: small hairpin. *: p<0.05; **: p<0.01 (two-way ANOVA).