Fig. 7: MacroD1 reduction inactivates NLRP3 inflammasome under LPS.
A Heatmap showing the expression of NLRP3 inflammasome pathway-related genes in the hearts of mice injected intraperitoneally with/without LPS (10 mg/kg) for 18 h based on bulk RNA sequence analysis. B–D Expression of IL-1β, IL-18, and TNF-α mRNAs in the hearts of mice with/without LPS administration (n = 5). E–H Expression of NLRP3 inflammasome pathway-related proteins in the heart of mice 18 h after intraperitoneal LPS (10 mg/kg) injection (n = 4). I–K Expression of ASC and NLRP3 proteins in the cytoplasm (Cyto; I and J) and mitochondria (Mito; I and K) isolated from hearts of mice together with LPS administration (n = 4). L Co-IP assay for ASC-NLRP3 interaction in the hearts of mice, together with LPS administration. M, N Proximity Ligation Assay fluorescence detecting interaction between ASC and NLRP3 in isolated ventricular cardiomyocytes from adult mice treated or untreated with LPS (1 μg/ml) for 18 h (n = 60). Red dots show ASC-NLRP3 interaction. Dotted lines outline cardiomyocytes. O–R Expression of NLRP3 pathway-related proteins in neonatal rat cardiomyocytes transfected with Si-MacroD1 (Si-M) or scrambled control (NC) for 36 h and administered ATP (3 mM) 18 h after LPS (1 μg/ml) treatment for 2 h (n = 4). S Representative fluorescence images examining co-localization of ASC (green), NLRP3 (red), and DAPI (blue) staining in NRCMs transfected with Si-MacroD1 (Si-M) or scrambled control (NC) for 36 h and administered ATP (3 mM) 18 h after saline or LPS (1 μg/ml) treatment for 2 h. Flox: MacroD1flox/flox controls; cKO: MacroD1flox/flox/αMHCMerCreMer mice. The three lines in the violin plot represent the third quartile, median, and first quartile, respectively, while other data are shown as mean ± SD. Data analysis was conducted utilizing one-way ANOVA with Tukey’s multiple comparison test (B–D, N). Data analysis was conducted using a two-tailed unpaired Student t test (F, G, J, K, P–R). Source data are provided as a Source Data file.
