Fig. 1

Components and activation of large yellow croaker NLRP3 inflammasome A: Caspase-1 processing (p10) in nigericin (40 μmol/L)-stimulated HEK293T cells overexpressing Flag-LcCaspase-1, with or without other inflammasome components, Flag-LcNLRP3, and/or HA-LcASC. B: Gasdermin E (cleaved N-term) processing in HEK293T cells overexpressing HA-LcGSDME, with or without Flag-LcNLRP3, Myc-LcASC, and/or Flag-LcCaspase-1, stimulated with nigericin (40 μmol/L) or not. C: Live cell imaging of pyroptosis in nigericin (40 μmol/L)-stimulated HEK293T cells overexpressing HA-LcGSDME, with or without other inflammasome components, using SYTOX Green for nucleic acid staining. Scale bars: 50 μm. D: Caspase-1 processing (p10) upon NLRP3 inflammasome activation in isolated large yellow croaker head kidney LcMO/MΦs primed with LPS and stimulated with nigericin or ATP for 60 min. E: Immunofluorescence images showing colocalization of LcNLRP3 with TGN38 (trans-Golgi network marker), TOMM20 (mitochondria marker), and calnexin (ER marker) in HEK293T cells overexpressing LcNLRP3 inflammasome components and stimulated with nigericin. Scale bars: 5 μm. F: Immunofluorescence images showing formation of LcNLRP3-ASC puncta and colocalization of NLRP3 puncta with MTOC marker GTU in HEK293T cells overexpressing LcNLRP3 inflammasome components, with or without nigericin stimulation. Scale bars: 5 μm. G: Immunofluorescence images showing colocalization of ASC puncta with MTOC marker GTU and colocalization of LcASC and LcCaspase-1 in HEK293T cells overexpressing LcNLRP3 inflammasome components, with or without nigericin stimulation. Scale bars: 5 μm. All immunofluorescence images are representative of three independent experiments. Blue indicates nuclear staining by DAPI. Arrowheads indicate puncta.