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: Frontiers in Immunology

Article Title: Persistent activation of monocytes/macrophages and cell senescence in SIV-infected macaques on ART

doi: 10.3389/fimmu.2026.1788994

Figure Lengend Snippet: NF-κB and Caspase-1 activation persists in spleens of SIV-infected RMs despite ART. (a) Representative Western blot analysis of phosphorylated NF-κB p65 (Ser536), total NF-κB p65, cleaved caspase-1 p20 and procaspase-1 in splenic lysates from uninfected control and SIV-infected RMs on ART. (b) Densitometric quantification of the phospho–NF-κB/total NF-κB and caspase-1 p20/procaspase-1 ratio is indicated. Data represent mean ± SEM; * P < 0.05 and ** P < 0.01 by unpaired t test.

Article Snippet: Primary antibodies were as follows: rabbit mAb against phospho-NF-κB p65 (Ser536) (93H1) (#3033, Cell Signaling Technology), NF-κB p65 (D14E12) XP ® (#8242, Cell Signaling Technology) and CASPASE 1 p20 (Cleaved Asp296) (PA5-99390, Invitrogen), and mouse mAb antibody against PROCASPASE-1 (D-3) (sc-392736, Santa Cruz Biotechnology).

Techniques: Activation Assay, Infection, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: The role of gasdermin-mediated mitochondrial RNA release in amplifying secondary immune response during microbial infection

doi: 10.3389/fimmu.2025.1668763

Figure Lengend Snippet: Gasdermin D and E mediated mtRNA release and VISA pathway activation induce secondary inflammatory response. Cells treated with LPS and Ng trigger the activation of the NLRP3 pathway, leading to the recruitment of caspase-1, which in turn cleaves GSDMD . In contrast, cells infected with VSV activate the PKR pathway and caspase-3, resulting in the cleavage of GSDME ( , ). Our findings indicate that the N-terminal fragments of both GSDMD and GSDME facilitate mitochondrial membrane permeabilization and the release of mtRNA into the cytoplasm. This mtRNA is subsequently recognized by the double-stranded RNA sensors RIG-I and MDA5, prompting their activation . Activated RIG-I interacts with VISA (MAVS) on the mitochondrial membrane , initiating a signaling cascade that involves various kinases, including TBK-1, IKKϵ, IKKα, and IKKβ ( , ). This cascade culminates in the phosphorylation and activation of key transcription factors, notably IRF-3/7 and NF-κB. These activated transcription factors then translocate to the nucleus to drive the expression of type I interferons (IFNs) and an array of pro-inflammatory cytokines ( , ). Furthermore, the N-terminal fragments of Gasdermin D and E contribute to the formation of pores in the plasma membrane, facilitating the release of mtRNA into the extracellular space due to the permeabilization induced by GSDMD and GSDME.

Article Snippet: Monoclonal rabbit antibodies against GSDME (ab215191) were purchased from Abcam at a dilution of 1:2000, and monoclonal rabbit antibodies against GSDMD (97558) from Cell Signaling Technology at a dilution of 1:1000, monoclonal rabbit antibodies against phospho-TBK1 (ab109272) from Abcam at a dilution of 1:2000, monoclonal rabbit antibody against Caspase-1 (3866) purchased from Cell Signaling Technology at a dilution of 1:1000, monoclonal rabbit antibody against NLRP3 (15101) sourced from Cell Signaling Technology at a dilution of 1:1000 and monoclonal antibody against VDAC1 (55259-1-AP) from Proteintech.

Techniques: Activation Assay, Infection, Membrane, Phospho-proteomics, Expressing, Clinical Proteomics

Journal: Frontiers in Immunology

Article Title: The role of gasdermin-mediated mitochondrial RNA release in amplifying secondary immune response during microbial infection

doi: 10.3389/fimmu.2025.1668763

Figure Lengend Snippet: GSDMD inhibition modulates cytokine production in LPS and Ng responses. (A, B) THP1 cells were primed with 100 ng/mL LPS for 2 hours and activated with 5 μM/mL nigericin (Ng) for one hour, with qPCR analysis demonstrating elevated expression of IFN-β (A) and IL-6 (B) . An unpaired t-test determined statistical significance. Data represent ± SEM (**p < 0.01, ***p < 0.001). (C, D) mouse peritoneal macrophages (PM cells) primed with 100 ng/mL LPS for 2 hours, treated with 5 μM/mL Ng for one hour, qPCR analysis demonstrating elevated expression of IFN-β (C) and IL-6 (D) , an unpaired t-test determined statistical significance. Data represent ± SEM (**p < 0.01, **p < 0.01). (E, F) THP-1 and PM cells primed with 100 ng/mL LPS alone exhibited reduced NLRP3 activation, caspase-1, and GSDMD cleavage. In contrast, cells primed with LPS and activated with 5 μM Ng showed enhanced NLRP3 activation, caspase-1, and GSDMD cleavage, leading to TBK1 phosphorylation. Cleaved caspase-1 protein (p20) was isolated from the supernatant (SN), and pTBK1 protein expression levels were quantified using ImageJ; an unpaired t-test determined statistical significance. Data represent ± SEM (***p < 0.001, ****p < 0.0001). (G, H) THP1 cells primed with 100 ng/mL LPS alone, cells treated with LPS + Ng, cells pretreated with 20 μM VX-765 for 2 hours and then treated with LPS + Ng, qPCR analysis showing decreased IFN-β (E) and IL-6 (F) expression in VX-765 pretreated THP-1 cells. Data represent mean ± SEM (*p < 0.05, ***p < 0.001). (I, J) PM cells primed with LPS alone, cells treated with LPS + Ng, cells pretreated with 20 μM VX-765 for 2 hours and then treated with LPS + Ng, qPCR analysis showing decreased IFN-β (G) and IL-6 (H) expression in VX-765 pretreated PM cells. Data represent mean ± SEM (**p < 0.01, *p < 0.05). (K, L) Western blot analysis of THP1 and PM cells showed no GSDMD cleavage and TBK1 phosphorylation in LPS-primed cells alone and cells pretreated with 20 μM VX-765 for 2 hours followed by LPS + Ng treatment. In contrast, LPS + Ng treatment induced GSDMD cleavage and elevated TBK1 phosphorylation, pTBK1 protein expression levels were quantified using ImageJ; an unpaired t-test determined statistical significance. Data represent ± SEM (****p < 0.0001, ***p < 0.001). (M, N) THP1 cells primed with LPS alone, cells primed with LPS, then treated with Ng, cells primed with LPS, then treated with 100 μM/mL dimethyl fumarate (DMF) for 2 hours and then treated with Ng, qPCR analysis showing decreased IFN-β (K) and IL-6 (L) expression in DMF treated THP1 cells. Data represent mean ± SEM (***p < 0.001, ****p < 0.0001). (O, P) PM cells primed with LPS alone, cells primed with LPS and then treated with Ng, cells primed with LPS, then treated with 100 μM/mL DMF for 2 hours and then treated with Ng, qPCR analysis showing decreased IFN-β (M) and IL-6 (N) expression in DMF treated PM cells. Data represent mean ± SEM (**p < 0.01, **p < 0.01). (Q, R) Western blot analysis showed that LPS priming, followed by Ng treatment, induced GSDMD cleavage and elevated TBK1 phosphorylation in THP-1 and PM cells. However, cells primed with LPS, treated with DMF, and treated with Ng inhibited GSDMD cleavage and reduced TBK1 phosphorylation, pTBK1 protein expression levels were quantified using ImageJ; an unpaired t-test determined statistical significance. Data represent ± SEM (****p < 0.0001, ***p < 0.001), all experiments were repeated three times, and representative experiments are shown.

Article Snippet: Monoclonal rabbit antibodies against GSDME (ab215191) were purchased from Abcam at a dilution of 1:2000, and monoclonal rabbit antibodies against GSDMD (97558) from Cell Signaling Technology at a dilution of 1:1000, monoclonal rabbit antibodies against phospho-TBK1 (ab109272) from Abcam at a dilution of 1:2000, monoclonal rabbit antibody against Caspase-1 (3866) purchased from Cell Signaling Technology at a dilution of 1:1000, monoclonal rabbit antibody against NLRP3 (15101) sourced from Cell Signaling Technology at a dilution of 1:1000 and monoclonal antibody against VDAC1 (55259-1-AP) from Proteintech.

Techniques: Inhibition, Expressing, Activation Assay, Phospho-proteomics, Isolation, Western Blot