NLRP3 Antibody - #AF4620

Figure 6 |TRPM2-shRNA-1 reduces OGD-induced expression of NLRP3,CXCL2, and caspase-1.(A)NLRP3 (NLRP3/β-actin), CXCL2 (CXCL2/β-actin), and caspase-1(caspase-1/β-actin) mRNA expression(as determined by fluorescence-based quantitative reverse transcription-polymerase chain reaction). (B) NLRP3, CXCL2, and caspase-1 protein expression (as determined by western blotting). Data are expressed as the mean ±standard deviation (SD) (n = 6; one-way analysis of variance followed by Student-Newman-Keuls post hoc test). *P< 0.05, vs. shRNA-NC group; #P < 0.05,vs. shRNA-NC + OGD group. CXCL2:chemokine ligand-2; NC: normal control; NLRP3: NACHT, LRR and PYD domains-containing protein 3; OGD:oxygen-glucose deprivation; TRPM2:transient receptor potential melastatin 2.

Fig. 6. Piezo1 on PANoptosis triggered by bone infection. (a) Infection human clinical tissues and healthy human clinical tissues: PIEZO1 and PANoptosis related markers (CASP1, CASPA3, NLRP3) were assessed through immunohistochemical staining. (b) Infected bone marrow cavity tissues and healthy tissues of rats: PIEZO1 and PANoptosis related markers (GSDMD, CASPA3, RIPK3) were assessed through immunohistochemical staining. (c) Semi-quantitative analysis of PIEZO1 and PANoptosis related markers (GSDME, CASPASE3, RIPK3) in the human clinical tissues. (d) Semi-quantitative analysis of PIEZO1 and PANoptosis related markers (GSDME, CASPASE3, RIPK3) in the bone marrow cavity tissue of rats. (e) Western blot of PANoptosis related markers (GSDMD, CASP1, CASP3, CASP8, pRIPK3, pMLKL). n = 3 independent experiments per group, *P < 0. 05; **P < 0. 01.

Fig 5. TGFBR1 gene silencing alleviates myocardial PANoptosis-like cell death in HFpEF mice. (A) Representative TUNEL staining images of the left-ventricular samples from each group of mice. Bar = 50 μm. (B) Quantification of TUNEL-positive cells in (A); n = 100 cells per group. (C) Representative western blot images of NLRP3, caspase-1, and GSDMD expression in the left-ventricular samples from each group of mice. (D) Representative western blot images of caspase-3, caspase-7, and caspase-8 expression in the left-ventricular samples from each group of mice. (E) Representative western blot images of p-RIPK1, p-RIPK3, and p-MLKL expression in the left-ventricular samples from each group of mice. (F)–(I) Quantification of the protein expression of NLRP3, cleaved caspase-1, and GSDMD-N in (C). (J)–(L) Quantification of the protein expression of cleaved caspase-3, cleaved caspase-7, and cleaved caspase-8 in (D). (M)–(O) Quantification of the protein expression of p-RIPK1, p-RIPK3, and p-MLKL in (E); n = 6 mice per group for panels A–O. The normality of data distribution was tested using the Shapiro–Wilk method. One-way ANOVA was applied in (B) and in (F)–(O). *p 

Fig. 3. The level of splenic CD4+ and CD8+ T lymphocytes and the expression of related proteins of NLRP3 inflammasome and Sirt1/autophagy axis in CLP rat. (X ±s, n=3). A. The CD4+ T cells in spleens of CLP rats by flow cytometry. B. The CD8+ T cells in spleens of CLP rats by flow cytometry. C. Bands of proteins; D-H. The statistical analysis results of NLRP3, Sirt1, Pro-IL-1β, Beclin-1, and LC3 Ⅱ/LC3 Ⅰ proteins bands by Western blot. Sham means the group underwent sham operation; CLP meats the group underwent CLP operation; HNK means the group was injected intraperitoneally with HNK 24h after the operation; And all rats were injected intraperitoneally with HNK (5 mg/kg twice) or PBS in the same volume. HNK: honokiol; CLP: cecal ligation and puncture; PBS: Phosphate buffer saline; NLRP3: NOD-like receptor thermal protein domain associated protein 3; Sirt1: Sirtuin1; ProIL-1β: Pro-Interleukin-1β; LC3: MAP1LC3.