FIGURE 5.

Gasdermin B (GSDMB) binds RNA as a potential RNA sensor. a) In vitro binding of biotin-high molecular weight (HMW) polyinosinic-polycytidylic acid (poly(I:C)) (3 µg·mL⁻¹) and immunoprecipitated Flag-GSDMB from BEAS-2B cells with stable expression of Flag-GSDMB detected by immunoprecipitation (IP) with streptavidin beads and immunoblot analysis with anti-Flag antibody. b, c) Dose-dependent competitive binding of unlabelled HMW poly(I:C) (b), but not unlabelled poly(deoxyadenylic-deoxythymidylic) acid (poly(dA:dT) (c) with Flag-GSDMB against biotin-labelled poly(I:C) (3 µg·mL⁻¹). d) Pure Flag-GSDMB instead of Flag-GSDMD protein binds to biotin-HMW poly(I:C) as shown by IP with neutravidin beads and immunoblotting analysis with anti-Flag antibody. e) Distribution of GSDMB electric charge was predicted based on each amino acid in the protein sequence. f) Haemagglutinin (HA)-GSDMB and its deletion mutants underwent IP and were eluted from BEAS-2B cells after transfection, followed by subsequent IP for their binding with biotin-labelled HMW poly(I:C). For all immunoblot data, representative results are shown from two independent biological experiments. His: histidine; P-W: position-window.