Fig 2: LCZ696 suppresses the activation of the NLRP3 inflammasome and inhibits the expression of proteins associated with the TAK1/JNK pathway in a rat model of MI. (A) Levels of TAK1 protein and phosphorylation of JNK (p-JNK) and p-JNK/JNK as detected by western blotting. (B) mRNA expression levels of TAK1 and JNK, as determined by reverse transcription-quantitative PCR. (C) Dual-fluorescent labelling staining of TAK1/NLRP3 (scale bar=50 µm). (D) Dual-fluorescent labelling staining of p-JNK/NLRP3 (scale bar=50 µm). Data are given as mean ± standard deviation, *P<0.05. LCZ696, sacubitril/valsartan; NLRP3, NLR pyrin family domain containing 3; TAK1, transforming growth factor ß-activated kinase-1; p-, phosphorylated.

Fig 3: ROS status and associated signalling molecules ERK1/2, JNK, and NRF-2 in the MDA-MB-231 cell line. ROS status indicators were the levels of (a) HIF-1a, (b) intracellular H2O2, (c) GSH, and (d) MDA; activities of (e) GPX, (f) CAT, and (g) SOD. ROS-sensing signalling molecules were (h) NRF-2 level; fold change in protein expression of (i) ERK1/2, (j) JNK, and (k) representative western blot analysis, SDS-PAGE of ERK1/2, JNK, and ß- actin. Results are expressed as the mean and standard error of the mean (SEM) (n = 3). Columns with dissimilar letters (a, b, c…) overhead within the same histogram are significantly different, and columns that have the same letters are not significantly different at p < 0.05. MDA group: untreated MDA-MB-231 cells served as control, MDA + Eb-ZFO group: MDA-MB-231 cells treated with Ebselen (Eb) and ZnFe2O4 nanoparticles (ZFO), MDA + IR group: MDA-MB-231 cells exposed to ?-radiation; and MDA + Eb-ZFO + IR group: MDA-MB-231 cells treated with Eb-ZFO and exposed to IR.

Fig 4: Activation of the TAK1/JNK pathway leads to accumulation of the NLRP3 inflammasome and the induction of cellular pyroptosis, resulting in increased inflammatory responses in H9c2 cells. (A) Relative expression levels of proteins, as detected by western blotting. (B) mRNA expression levels of TAK1, JNK, NLRP3, caspase-1 and GSDMD, as determined by reverse transcription-quantitative PCR. (C) Levels of IL-1ß and IL-18, in samples of rat serum, as detected by ELISA. Data were given as mean ± standard deviation, *P<0.05. TAK1, transforming growth factor ß-activated kinase-1; NLRP3, NLR pyrin family domain containing 3; GSDMD, gasdermin D; p-, phosphorylated.

Fig 5: Effect of compound 1 on the phosphorylation of MAPK, (A) JNK, (B) p38, (C) ERK and (D) STAT3 in RAW 264.7 cells. All data are presented as the mean± SEM (n = 3). Quercetin (Q5, 5 µg/mL) was used as the positive control. * p < 0.05 and ** p < 0.01, *** p < 0.001 indicate significant differences compared to the LPS control group.