caspase recruitment domain  (Cell Signaling Technology Inc)

Journal: Liver Research

Article Title: Aluminum adjuvant promotes liver inflammation and fibrosis in mice: A novel approach to establish a liver fibrosis animal model

doi: 10.1016/j.livres.2025.05.001

Figure Lengend Snippet: Imject Alum increased the level of pro-inflammatory factors and inflammatory cells in the liver. (A) qRT-PCR analysis of specific leukocyte markers in the liver of Imject Alum (Al(OH) 3 160 mg/kg) D54 and control (CTR) groups. CTR group, n = 8; Imject Alum (Al(OH) 3 160 mg/kg) D54 group, n = 10. ∗ P < 0.05, compared with the CTR group. ns, no significant. (B) Representative immunohistochemical staining with F4/80 for macrophages in the liver from different groups. (C) Representative immunohistochemical staining with Cd3e for T lymphocytes in the liver from different groups. (D) Representative immunofluorescence staining of hepatic Cd11b. (E) Quantitative analysis of hepatic F4/80 and Cd3e immunohistochemical staining. CTR group, n = 8; Imject Alum (Al(OH) 3 160 mg/kg) D54 group, n = 10. ∗ P < 0.05, compared with the CTR group. (F) Hepatic mRNA expression of cytokines, chemokines, and cell adhesion molecules (CAMs) significantly increased in Imject Alum (Al(OH) 3 160 mg/kg) D54 group. CTR group, n = 8; Imject Alum (Al(OH) 3 160 mg/kg) D54 group, n = 10. ∗ P < 0.05, compared with the CTR group. (G) Relative levels of pyroptosis protein markers in the CTR and Imject Alum (Al(OH) 3 160 mg/kg) D54 groups. (H) Primary mouse hepatocyte mRNA levels of pyroptosis markers were significantly increased after treating with Imject Alum. n = 3. ∗ P < 0.05, compared with PBS group. # P < 0.05, compared with Imject Alum (Al(OH) 3 0.5 mg/mL) group. (I) The NLRP3 inhibitor MCC950 can inhibit the activation of primary mouse hepatocyte pyroptosis induced by Imject Alum. n = 3. ∗ P < 0.05, compared with PBS group. # P < 0.05, compared with Imject Alum (Al(OH) 3 1 mg/mL) group. Abbreviations: Asc, apoptosis-associated speck-like protein containing a caspase recruitment domain; Ccl2, C–C motif ligand 2; Cxcl, CXC motif chemokine ligand; Gsdmd, gasdermin D; Icam1, intercellular adhesion molecule 1; Il, interleukin; MOD, mean optical density; Nlrp3, nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain-containing 3; PBS, phosphate-buffered saline; qRT-PCR, quantitative real-time polymerase chain reaction; Tnf-α, tumor necrosis factor-alpha; Vcam1, vascular cell adhesion molecule 1.

Article Snippet: The primary antibodies used in this study were anti-Gapdh (1:10000 dilution, #A19056, ABclonal, Wuhan, China); anti-Nlrp3 (1:1000 dilution, #15101S, Cell Signaling Technology, Danvers, MA, USA); anti-Caspase 1 (1:1000 dilution, #22915-1-AP, Proteintech, Wuhan, China); anti-gasdermin D (anti-Gsdmd; 1:1000 dilution, #AF4012, Affinity, Changzhou, Jiangsu, China); and anti-apoptosis-associated speck-like protein containing a caspase recruitment domain (anti-Asc; 1:1000 dilution, #67824T, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Quantitative RT-PCR, Control, Immunohistochemical staining, Staining, Immunofluorescence, Expressing, Activation Assay, Binding Assay, Saline, Real-time Polymerase Chain Reaction

Journal: Translational stroke research

Article Title: Catecholamine-Induced Inflammasome Activation in the Heart Following Photothrombotic Stroke.

doi: 10.1007/s12975-024-01311-3

Figure Lengend Snippet: Fig. 3 Caspase-1 and IL-1β are reduced in the heart following IC100 administration after PTS. A Representative immunoblots of atria lysates and quantifi- cation indicate significantly reduced levels of caspase-1 in mice injected with IC100 24 h post-PTS. B 24 h post-PTS, cas- pase-1 is statistically reduced in the atria of IC100-treated mice compared to saline-treated mice. Data shown as mean + / − SEM. p-values shown for Student’s T test. N = 4 to 5 per group. C IC100 decreased IL-1β in the atria after PTS. D Representa- tive immunoblots of ventricle lysates and quantification show- ing no difference between saline or IC100 treated mice 24 h post PTS. E 24-h post-PTS, the expression of caspase-1 did not differ in the ventricle compared to saline-treated mice. F Ven- tricle lysate expression of IL-1β was reduced in in IC100-treated mice 24 h post-PTS. (B) Data shown as mean + / − SEM. N = 4 to 6 per group

Article Snippet: For immunoblot analysis of inflammasome signaling proteins, protein lysates were resolved in 4–20% Criterion TGX Stain-Free precast gels (Bio-Rad), using antibodies (1:1000 dilution) against AIM2 (Novus), IL-1β (1:500, Cell Signaling), apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC) (Santa Cruz), Caspase-1 (Novus Biological), Caspase-8 (Novus Biological) as described in [41].

Techniques: Western Blot, Injection, Saline, Expressing