Journal: Acta Biochimica et Biophysica Sinica

Article Title: Selective inhibition of the NLRP3 inflammasome protects against acute ethanol-induced cardiotoxicity in an FBXL2-dependent manner

doi: 10.3724/abbs.2023256

Figure Lengend Snippet: Influence of MCC950 on acute ethanol exposure-induced changes in FBXL2 and NLRP3 levels and effect of NLRP3 silencing or AC-YVAD-CMK on acute ethanol exposure-induced changes in cleaved caspase-1 and FBXL2 (A) Representative immunoblots depicting levels of FBXL2 using specific antibody in control and ethanol mice treated with or without MCC950 (20 μM). (B) Pooled FBXL2 level. (C) Representative immunoblots depicting levels of NLRP3, cleaved caspase-1, and FBXL2 using specific antibodies in control and ethanol-challenged cells with or without NLRP3 silencing. (D) Pooled NLRP3 level. (E) Cleaved caspase-1 level. (F) Pooled FBXL2 level. (G) Representative immunoblots depicting levels of cleaved caspase-1 and FBXL2 using specific antibodies in control and ethanol-challenged cells with or without AC-YVAD-CMK (50 μM) treatment. (H) Pooled cleaved caspase-1 level. (I) Pooled FBXL2 level. Data are presented as the mean±SEM. n=6 for panel B and n=3 for the remaining panels. *P<0.05 vs control (CON) group; #P<0.05 vs ethanol group.

Article Snippet: Drugs such as MCC950 (Sigma-Aldrich, St Louis, USA), caspase-1 inhibitor AC-YVAD-CMK (Sigma-Aldrich), FBXL2 activator BC-1258 (Sigma-Aldrich), and FBXL2 inhibitor GGTI-2418 (Med Chem Express, Princeton, USA) were applied to mice or cardiomyocytes.

Techniques: Western Blot, Control

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Selective inhibition of the NLRP3 inflammasome protects against acute ethanol-induced cardiotoxicity in an FBXL2-dependent manner

doi: 10.3724/abbs.2023256

Figure Lengend Snippet: Influence of FBXL2 activation and FBXL2 colocalization inhibition on MCC950-mediated protection against acute ethanol exposure-induced defects in cardiomyocyte contractile function Adult murine cardiomyocytes were exposed to ethanol (160 mM) in the presence or absence of the FBXL2 activator BC-1258 (10 μM) or the FBXL2 colocalization inhibitor GGTi-2418 (10 μM) prior to assessment of cardiomyocyte function. (A) Resting cell length. (B) Peak shortening (normalized to cell length). (C) Maximal velocity of shortening (+dL/dt). (D) Maximal velocity of relengthening (–dL/dt). (E) Time-to-peak shortening (TPS). (F) Time to 90% relengthening (TR90). n=37 cells from 2 mice/group. Data are presented as the mean±SEM. *P<0.05 vs control (CON) group; # P<0.05 vs ethanol group; †P<0.05 vs EtOH-MCC950 group.

Article Snippet: Drugs such as MCC950 (Sigma-Aldrich, St Louis, USA), caspase-1 inhibitor AC-YVAD-CMK (Sigma-Aldrich), FBXL2 activator BC-1258 (Sigma-Aldrich), and FBXL2 inhibitor GGTI-2418 (Med Chem Express, Princeton, USA) were applied to mice or cardiomyocytes.

Techniques: Activation Assay, Inhibition, Control

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Selective inhibition of the NLRP3 inflammasome protects against acute ethanol-induced cardiotoxicity in an FBXL2-dependent manner

doi: 10.3724/abbs.2023256

Figure Lengend Snippet: Influence of FBXL2 activation and FBXL2 colocalization inhibition on NLRP3 silencing-offered protection against acute ethanol exposure-induced changes in mitochondrial membrane potential and mitochondrial LC3B puncta Neonatal rat cardiomyocytes were exposed to ethanol (160 mM) in the presence or absence of the FBXL2 activator BC-1258 (10 μM) or the FBXL2 colocalization inhibitor GGTi-2418 (10 μM) prior to assessment of mitochondrial membrane potential and LC3B puncta formation and colocalization in mitochondria (using COXIV as a marker). (A) Representative JC-1 fluorescence images depicting the effect of BC-1258 and GGTi-2418 on NLRP3 silencing-mediated protection against acute ethanol exposure-induced collapse of mitochondrial membrane potential. CCCP was used as a positive control. Scale bar: 100 μm. (B) Representative LC3B puncta and COXIV fluorescence images depicting the effect of BC-1258 and GGTi-2418 on NLRP3 silencing-offered protection against acute ethanol exposure-induced mitochondrial autophagosome formation. Scale bar: 10 μm. (C) Pooled data of JC-1 using the aggregate-to-monomer ratio. (D) Pooled data depicting colocalization of LC3B and COXIV. n=6. Data are presented as the mean±SEM. *P<0.05 vs control (CON) group; # P<0.05 vs ethanol group; †P<0.05 vs EtOH-siNLRP3 group.

Article Snippet: Drugs such as MCC950 (Sigma-Aldrich, St Louis, USA), caspase-1 inhibitor AC-YVAD-CMK (Sigma-Aldrich), FBXL2 activator BC-1258 (Sigma-Aldrich), and FBXL2 inhibitor GGTI-2418 (Med Chem Express, Princeton, USA) were applied to mice or cardiomyocytes.

Techniques: Activation Assay, Inhibition, Membrane, Marker, Fluorescence, Positive Control, Control