Journal: Oxidative Medicine and Cellular Longevity

Article Title: Recurrent Hypoglycemia Impaired Vascular Function in Advanced T2DM Rats by Inducing Pyroptosis

doi: 10.1155/2022/7812407

Figure Lengend Snippet: NF- κ B, NLRP3, and cGAS–STING pathway activity of the aortas in response to acute and recurrent hypoglycemia in aged T2DM rats. (a) p-p65, NLRP3, ASC, (b) cleaved caspase-1, cGAS, and (e) STING expression were assayed by western blotting. The expression and location of NLRP3 were determined by (c) immunohistochemistry and (d) immunofluorescence; ∗ p < 0.05 DM vs. control; # p < 0.05 H-DM, RH-DM vs. DM; & p < 0.05 H-DM vs. RH-DM.

Article Snippet: Proteins were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis and incubated with primary antibodies: eNOS (1 : 1000, ab300071, Abcam), iNOS (1 : 1000, ab283655, Abcam), NOX2 (1 : 2000, 19013-1-AP, ProteinTech), NOX4 (1 : 2000, 14347-1-AP, ProteinTech), p-p65 (1 : 1000, 3033, Cell Signaling Technology), p65 (1 : 1000, 8242, Cell Signaling Technology), NLRP3 (1 : 1000, NBP2-12446, NOVUS), ASC (1 : 1000, sc-514414, Santa Cruz), Caspase-1 (1 : 1000, bs-10743R, Bioss), cGAS (1 : 1000, NBP3-16666, NOVUS), STING (1 : 1000, CST50494, Cell Signaling Technology), GSDMD (1 : 1000, NBP2-33422, NOVUS), Bax (1 : 1000, 50599-2-Ig, ProteinTech), Bcl-2 (1 : 1000, 26593-1-AP, ProteinTech), and β -actin (1 : 5000, 20536-1-AP, ProteinTech).

Techniques: Activity Assay, Expressing, Western Blot, Immunohistochemistry, Immunofluorescence

Journal: Journal of Nanobiotechnology

Article Title: Manganese-enriched nanoboron agent amplifies BNCT efficacy via pyroptosis-mediated immune activation and STING pathway synergy​

doi: 10.1186/s12951-025-03916-8

Figure Lengend Snippet: Pyroptosis Induced by BSA- 10 BPA-MnO 2 after BNCT. ( a ) Fluorescent images of ROS in B16F10 cells after different treatments, detected by DCFH-DA staining. Scale bar: 150 μm. ( b ) Cell morphology images after different treatments. In the BSA- 10 BPA-MnO 2 + N group, cell swelling and the appearance of large bubbles (blue arrows, one of the most prominent features of pyroptosis) are observed. Scale bar: 50 μm. ( c ) Western blot analysis of caspase-1 (CAS-1) and cleaved caspase-1 (cleaved-CAS-1). ( d ) Immunofluorescent staining of the GSDMD protein N-terminal fragment (GSDMD-N). Scale bar: 100 μm. ( e ) Relative LDH release, n = 4. Data are presented as mean ± SD. * p < 0.5, **** p < 0.0001

Article Snippet: After blocking, membranes were probed with primary antibodies against caspase-1 and cleaved caspase-1 (MedChemExpress, China), followed by HRP-conjugated secondary antibodies.

Techniques: Staining, Western Blot

Journal: Science advances

Article Title: Targeting pyroptosis with nanoparticles to alleviate neuroinflammatory for preventing secondary damage following traumatic brain injury.

doi: 10.1126/sciadv.adj4260

Figure Lengend Snippet: Fig. 4. Inhibitory effect of C-β-LG/DSF on pyroptosis after TBI. (A) The schematic diagram illustrates the inhibition of pyroptosis by DSF. After the uptake of C-β-LG/DSF by cells, DSF inhibits the aggregation of GSDMD N-terminal pores on the cell membrane surface. Furthermore, experimental results demonstrated that the expression of GSDMD, GSDMD-N, caspase-1, and other related proteins was reduced after treatment with C-β-LG/DSF. (B) Immunoblotting of GSDMD, GSDMD-N, and caspase-1, caspase-1 p10, caspase-1 p20 in neuron-ICR cells with sham, TBI, DSF, and C-β-LG/DSF for 24 hours. (C to E) Levels of (C) IL-1β, (D) IL-18, and (E) LDH in injured neuron-ICR as detected by ELISA 12 hours after TBI. (F) Immunohistochemical staining was used to observe the expression of GSDMD in injured tissues of TBI model mice 3 days after different drugs treatment. Scale bars, 50 μm. (G) Quantitative analysis of GSDMD-positive cells. (H to J) Levels of (H) IL-1β, (I) IL-18, and (J) LDH in injured tissues as detected by ELISA 3 days after TBI. (K) Immunoblotting of GSDMD, GSDMD-N, caspase-1, caspase-1 p20, and caspase-1 p10 in injured tissue with sham, TBI, DSF, β-LG/DSF, and C-β-LG/ DSF for 24 hours. Data were expressed as means ± SD (n = 5). For (C) to (E) and (G) to (J), statistical analysis was calculated via one-way ANOVA test. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: Primary antibodies used for immunoblot were listed as follows: anti– caspase- 1 p20 (Santa Cruz Biotechnology, sc- 398715; 1:1000), anti– cleaved GSDMD (Cell Signaling Technology, #10137S; 1:1000), anti–caspase- 1 p10 (GeneTex, GTX134551; 1:1000), and anti- GAPDH (Proteintech, 60004- 1- Ig; 1:1000).

Techniques: Inhibition, Membrane, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining

Journal: Cell Proliferation

Article Title: Combination of resolvin E1 and lipoxin A4 promotes the resolution of pulpitis by inhibiting NF‐κB activation through upregulating sirtuin 7 in dental pulp fibroblasts

doi: 10.1111/cpr.13227

Figure Lengend Snippet: Primers used for the selected human genes

Article Snippet: Anti‐NLRP3, anti‐caspase1, anti‐IL‐1β, anti‐IL‐18 (mentioned above) and anti‐OCN (PB1009, BOSTER) antibodies were used as primary antibodies at 1:200 dilutions.

Techniques:

Journal: Experimental and Therapeutic Medicine

Article Title: 2,3,5,4′-tetrahydroxy-stilbene-2-O-β-D-glucoside attenuates methionine and choline-deficient diet-induced non-alcoholic fatty liver disease

doi: 10.3892/etm.2018.6300

Figure Lengend Snippet: The effects of TSG on protein expression of NLRP3, ASC and caspase-1 in livers of MCD diet fed mice. Protein imprinting of NLRP3, ASC, and caspase-1. (A) As a matter of fact, nine samples were included in the original gel/experiment. The antepenultimate and penultimate pair were omitted from the figure (as denoted by the dotted lines) since they were not relevant for a discussion of the experiment at hand. Densitometric analysis of (B) NLRP3, (C) ASC, (D) caspase-1. Mice fed an MCD diet showed increased levels of NLRP3, ASC and caspase-1 protein expressions compared with mice in the control diet-fed group. The results showed that the middle dosage of TSG downregulated ASC and caspase-1 levels. ## P<0.01; ### P<0.001 ( # , compared with MCD diet-induced NAFLD model group). *P<0.05; **P<0.01; ***P<0.001 (*, compared with the control group). TSG, 2,3,5,4′-tetrahydroxy-stilbene-2-O-β-D-glucoside; NLRP3, Nod-like receptor protein 3; ASC, apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain; MCD, methionine and choline-deficient; NAFLD, Non-alcoholic fatty liver disease.

Article Snippet: Antibodies directed against NLRP3, apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain (ASC) and caspase-1 were purchased from Proteintech Group, Inc. (Chicago, IL, USA).

Techniques: Expressing