anti human il 1β  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Microbiome-derived hydroxyphenyl propanoates enhance antitumour immunity by potentiating gasdermin D activity in tumour-associated myeloid cells

doi: 10.64898/2026.04.23.720410

Figure Lengend Snippet: a, IL-1β activity in the supernatant of THP1-WT vs. - GSDMD -KO cells treated with LPS±HPPs for 16 hours, measured using HEK-Blue™ IL-1β reporter cells (three combined experiments, error bars represent standard error, one-way ANOVA with Bonferroni test); b, LDH release from THP1-WT vs. – GSDMD -KO cells treated with LPS±HPP for 16 hours (three combined experiments, one-way ANOVA with Bonferroni test); c-d, IL-1β activity in the supernatant of THP1-WT vs. - GSDMD -KO cells © and human PBMCs (d) treated with LPS and NG±HPPs for 16 hours. measured using HEK-Blue™ IL-1β reporter cells (three combined experiments, error bars represent standard error, one-way ANOVA with Bonferroni test); e-f, LDH release from THP1-WT vs. – GSDMD -KO cells € and human PBMCs (f) treated with LPS and NG±HPPs for 16 hours (three combined experiments, one-way ANOVA with Bonferroni test).

Article Snippet: To neutralize IL-1 receptor signalling, the following antibodies were used: anti-human IL-1α (1 μg/mL; clone 7D4; mabg-hil1a-3; InvivoGen), anti-human IL-1β (1 μg/mL; clone 4H5; mabg-hil1b-3; InvivoGen) and IgG1 isotype control (1 μg/mL; clone T8E5; mabg1-ctrlm; InvivoGen).

Techniques: Activity Assay

Journal: bioRxiv

Article Title: Microbiome-derived hydroxyphenyl propanoates enhance antitumour immunity by potentiating gasdermin D activity in tumour-associated myeloid cells

doi: 10.64898/2026.04.23.720410

Figure Lengend Snippet: a-b, tumour growth kinetics and OS for orthotopic M3-9-M OVA tumours in male WT or Gsdmd -KO mice receiving vehicle or 3,2-HPP treatments from day 1 post cell-implantation (n =8/group; two combined experiments, two-way ANOVA test for growth kinetics, Log-Rank test for OS); c, [IL-1β] in interstitial fluid harvested from 18-day old M3-9-M OVA tumours grown in male WT or Gsdmd -KO mice receiving vehicle or 3,2-HPP treatment from day 1 post-cell implantation (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); d, GSDMD peptides quantified from a publicly available tumour proteomic database established from stage IV melanoma patients undergoing anti-PD-1 treatment (responder, R = 40 and non-responder, NR = 27); e-g, correlation uncleaved GSDMD peptide with NFKB1 gene targets and two sets of M1- vs. M2-like macrophage gene signatures, respectively, in stage IV melanoma patients undergoing anti-PD-1 treatment (R = 18 and NR = 6, chi-square test).

Article Snippet: To neutralize IL-1 receptor signalling, the following antibodies were used: anti-human IL-1α (1 μg/mL; clone 7D4; mabg-hil1a-3; InvivoGen), anti-human IL-1β (1 μg/mL; clone 4H5; mabg-hil1b-3; InvivoGen) and IgG1 isotype control (1 μg/mL; clone T8E5; mabg1-ctrlm; InvivoGen).

Techniques: Comparison

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH inhibits activation of the NLRP3 inflammasome. (A) Schematic workflow of in vitro NLRP3 inflammasome activation. (B,C) Cell viability was assessed in J774A.1 cells (B) and PMA (500 nM)-differentiated THP-1 cells (C) treated with TMH for 2 or 18 h using the EZ-Cytox assay. (D) LPS-primed J774A.1 cells were treated with TMH for 2 h, then stimulated with ATP (5 mM) for 30 min in the presence or absence of MCC950. IL-1β (p17) levels in supernatants were measured by ELISA. (E,F) PMA-differentiated THP-1 cells were primed with LPS, treated with TMH for 2 h, then stimulated with ATP (5 mM) (E) or nigericin (10 μM) (F) for 1 h, with or without MCC950. IL-1β (p17) levels in supernatants were determined by ELISA. Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; n.s., not significant.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Activation Assay, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH does not affect other inflammasome or inflammatory pathways. (A,B) LPS-primed J774A.1 cells were treated with TMH for 2 h, followed by stimulation with either dsDNA (1 μg/mL) (A) or flagellin (B) transfected using Lipofectamine 2000 for 3 h. (C) Following TMH treatment (2 h), nuclear and cytosolic fractions of LPS-primed J774A.1 cells were separated and assessed for NF-κB p65 translocation by immunoblotting. (D) HEK293FT cells transfected with the pGL4.32 luciferase reporter vector were treated with TNF-α (20 ng/mL) for 5 h, with or without TMH pre-treatment (2 h). Luciferase activity was measured using the Bright-Glo™ luciferase assay system (Promega). Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test n.s., not significant.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Transfection, Translocation Assay, Western Blot, Luciferase, Plasmid Preparation, Activity Assay

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH inhibits NLRP3 inflammasome activation by suppressing ASC oligomerization and ATPase activity. (A) LPS-primed J774A.1 cells were treated with TMH for 2 h, then stimulated with imiquimod (200 μM) for 1 h. (B) Cells were treated with TMH (2 h), stained with MitoSOX (5 μM, 10 min), and stimulated with ATP (5 mM, 5 min). Intracellular ROS were visualized by confocal microscopy. (C,D) PMA-differentiated THP-1 cells were primed with LPS, treated with TMH for 2 h, then stimulated with nigericin (10 μM, 1 h). ASC speck formation was examined by confocal microscopy (C) , and representative images from five fields are shown (D) . (E) Cells were stimulated with or without MCC950 and cross-linked with DSS (2.5 mM, 30 min) before immunoblotting for ASC oligomerization. (F) NLRP3 ATPase activity was measured using the ADP-Glo™ assay to quantify ATP-to-ADP conversion following TMH treatment. Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. **P < 0.01, ***P < 0.001.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Activation Assay, Activity Assay, Staining, Confocal Microscopy, Western Blot, Glo Assay

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: Toxicity assessment of TMH in zebrafish embryos. (A) Survival curves of zebrafish embryos treated with TMH at concentrations of 1, 10, or 100 μg/mL. Sixty embryos were used per condition and maintained in E3 medium. The control group was treated with DMSO (0.25%, v/v), corresponding to the highest TMH concentration. (B) Survival and phenotype distribution of 5 dpf embryos treated with TMH at distinct concentrations from 1 to 5 dpf. (C) Representative images showing developmental abnormalities observed in embryos treated with 10 μg/mL TMH at 5 dpf. (D) Representative images of Sudan Black B staining in 3 dpf embryos treated with vehicle control (0.0025% DMSO with PTU; upper panel) or TMH (1 μg/mL; lower panel). (E) Quantification of neutrophil counts in the CHT (red dashed outline) following treatment with TMH (1 μg/mL). Data are presented as mean ± SEM of individual embryos. Statistical significance was determined using an unpaired two-tailed Student’s t-test; n.s., not significant (p = 0.9997). Scale bars, 300 µm.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Control, Concentration Assay, Staining, Two Tailed Test

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH suppresses LPS-induced inflammatory cell recruitment in zebrafish embryos. (A) Representative images of neutrophil recruitment in the CHT in 3 dpf zebrafish embryos following LPS stimulation, assessed by Sudan Black B staining. Top panel: control embryos treated with 0.0025% DMSO (n = 29); middle panel: LPS-treated embryos (10 μg/mL; n = 32); bottom panel: embryos co-treated with LPS (10 μg/mL) and TMH (1 μg/mL; n = 33). (B) Quantification of neutrophil accumulation in the CHT following LPS stimulation with or without TMH treatment using Sudan Black B staining from (A) . (C) Representative images of whole-mount in situ hybridization showing mpx -positive neutrophils in 3 dpf embryos under LPS and TMH treatment conditions. Top panel (DMSO 0.0025%; n = 34); middle panel (LPS 10 μg/mL; n = 35); bottom panel (LPS 10 μg/mL with TMH 1 μg/mL; n = 39). (D) Quantification of mpx -positive cells within the CHT from (C) . (E) Representative confocal images of macrophages in 3 dpf Tg ( mpeg1 :EGFP) zebrafish embryos following LPS and TMH treatment. Top panel (DMSO 0.0025%; n = 8); middle panel (LPS 10 μg/mL; n = 11); bottom panel (LPS 10 μg/mL with TMH 1 μg/mL; n = 13). (F) Quantification of mpeg1 :EGFP-positive macrophages in the CHT region shown in (E) . Red dashed outline indicates the CHT. All data are presented as mean ± SEM. Statistical significance was determined using an unpaired two-tailed Student’s t-test. *P < 0.05; **P < 0.01; ***P < 0.05; ****P < 0.0001. Scale bars, 300 µm.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Staining, Control, In Situ Hybridization, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Investigating the impact and mechanism of Licochalcone B derivative CTG12 on NLRP3 inflammasome

doi: 10.1186/s12964-026-02741-2

Figure Lengend Snippet: CTG12 inhibits NLRP3 inflammasome activation in mouse BMDMs and human THP1. A The structure of CTG12. B , C Cell viability of BMDMs ( B ) and THP1 cells ( C ) treated with CTG12. (D-H) Western blot analysis of IL-1β (p17), caspase-1 (p20) in culture supernatants (SN) and pro-IL-1β, caspase-1 (p45), NLRP3, ASC in whole cell lysates (WCL) of LPS-primed BMDMs treated with CTG12 and then stimulated with Nigericin (25 min) ( D ), supernatants were collected for the measurement of caspase-1( E ), IL-1β ( F ), LDH ( G ) and TNF-α ( H ). (I-M) THP-1 cells primed with PMA for 12 h were treated with CTG12 for 1 h and then stimulated with nigericin for 55 min. Western blot analysis of IL-1β (p17), caspase-1 (p20) in culture supernatants (SN) and pro-IL-1β, caspase-1 (p45), ASC in whole cell lysates (WCL) of THP-1 ( I ), supernatants were collected for the measurement of caspase-1( J ), IL-1β ( K ), LDH ( L ) and TNF-α ( M ). Coomassie blue-stained gels was used as loading control and Lamin B was used as a control for equal loading of the samples. Data represent as mean ± SEM ( n = 3). Compared to con, ** p < 0.01, **** p < 0.0001; compared to a concentration of 0 μM, # p < 0.05, ## p < 0.01, #### p < 0.0001 and ns: not significant (one-way ANOVA with Dunnett’s post-hoc test)

Article Snippet: Anti-human IL-1β (p17, 12242S), anti-Phospho-IRF3 (Ser396) (4947), caspase-1 antibody (4199S) and anti-ASC antibody (sc-22, 514-R) were obtained from Cell Signaling Technology (Germany).

Techniques: Activation Assay, Western Blot, Staining, Control, Concentration Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Investigating the impact and mechanism of Licochalcone B derivative CTG12 on NLRP3 inflammasome

doi: 10.1186/s12964-026-02741-2

Figure Lengend Snippet: CTG12 specifically inhibits canonical and noncanonical NLRP3 inflammasome activation. A - C BMDMs were primed with LPS and then treated with CTG12 (5 μM), then stimulated with Nigericin, ATP, poly (I:C), or SiO₂. Western blot analysis of caspase-1 (p20) in culture supernatants (SN) and pro-IL-1β, pro-caspase-1 (p45), NLRP3 and ASC in whole-cell lysates (WCL); secretion of activated caspase-1 (p20) in the culture supernatant (SN) of BMDMs (A), supernatants were collected for the measurement of caspase-1 ( B ) and IL-1β ( C ). D - F BMDMs primed with Pam3CSK4 treated with CTG12 (5 μM), followed by cytosolic LPS. Western blot analysis of caspase-1 (p20) in culture supernatants (SN) and pro-IL-1β, pro-caspase-1 (p45), NLRP3 and ASC in whole-cell lysates (WCL); secretion of activated caspase-1 (p20) in the culture supernatant (SN) of BMDMs ( D ), supernatants were collected for the measurement of caspase-1 ( E ) and IL-1β ( F ). ( G - I ) BMDMs were primed with LPS treated with CTG12 (5 μM), then stimulated with Nigericin, Salmonella, poly (dA:dT). Western blot analysis of caspase-1 (p20) in culture supernatants (SN) and pro-IL-1β, pro-caspase-1 (p45), NLRP3 and ASC in whole-cell lysates (WCL); secretion of activated caspase-1 (p20) in the culture supernatant (SN) of BMDMs ( G ), supernatants were collected for the measurement of caspase-1 ( H ) and IL-1β ( I ). Coomassie blue-stained gels used as loading control and Lamin B used as a control for equal loading of the samples. Data represent as mean ± SEM ( n = 3). # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 and ns: not significant (one-way ANOVA with Dunnett’s post-hoc test)

Article Snippet: Anti-human IL-1β (p17, 12242S), anti-Phospho-IRF3 (Ser396) (4947), caspase-1 antibody (4199S) and anti-ASC antibody (sc-22, 514-R) were obtained from Cell Signaling Technology (Germany).

Techniques: Activation Assay, Western Blot, Staining, Control

Journal: Cell Communication and Signaling : CCS

Article Title: Investigating the impact and mechanism of Licochalcone B derivative CTG12 on NLRP3 inflammasome

doi: 10.1186/s12964-026-02741-2

Figure Lengend Snippet: CTG12 inhibits LPS-induced priming step of inflammasome activation and specifically inhibits canonical and noncanonical NLRP3 inflammasome activation by blocking NLRP3-dependent ASC oligomerization. A BMDMs were cultured with LPS for 4 h, then incubated with CTG12 for 1 h, or BMDMs were first treated with CTG12 for 1 h, followed by stimulation with LPS for 4 h, western blot analysis of protein levels of NLRP3, ASC, pro-IL-1β, Cas-1 p45, Lamin B. B BMDMs were primed with LPS treated with CTG12 (5 μM), then stimulated with Nigericin, western blot analysis of Triton X-insoluble microparticle cross-linking ASC. C BMDMs were primed with LPS treated with CTG12 (5 μM), then stimulated with Nigericin, ATP, poly(I:C), SiO 2 , western blot analysis of in Triton X insoluble microparticle cross-linked ASC. D BMDMs were primed with Pam3CSK4 treated with CTG12 (5 μM), followed by stimulation with ultra-LPS, western blot analysis of Triton X-insoluble microparticle cross-linking ASC. E BMDMs were primed with LPS treated with CTG12 (5 μM), then stimulated with Nigericin, salmonella or poly (dA:dT), western blot analysis of in Triton X insoluble microparticle cross-linked ASC. Coomassie blue-stained gels used as loading control and Lamin B used as a control for equal loading of the samples

Article Snippet: Anti-human IL-1β (p17, 12242S), anti-Phospho-IRF3 (Ser396) (4947), caspase-1 antibody (4199S) and anti-ASC antibody (sc-22, 514-R) were obtained from Cell Signaling Technology (Germany).

Techniques: Activation Assay, Blocking Assay, Cell Culture, Incubation, Western Blot, Staining, Control

Journal: Cell Communication and Signaling : CCS

Article Title: Investigating the impact and mechanism of Licochalcone B derivative CTG12 on NLRP3 inflammasome

doi: 10.1186/s12964-026-02741-2

Figure Lengend Snippet: CTG12 alleviates LPS-induced acute systemic inflammation in mice. A - J Female mice (8 weeks) were injected intraperitoneally with CTG12 (0 mg/kg, 15 mg/kg, 30 mg/kg), MCC950 (40 mg/kg) or CTG12 (30 mg/kg) + MCC950 (40 mg/kg) for 1 h, followed by an injection of 20 mg/kg LPS for 5 h ( n = 6). The levels of IL-1β (A, B), IL-18 ( C , D ), IL-6 ( E , F ), TNF-α ( G , H ) and CXCL1/KC ( I , J ) in peritoneal lavage fluid and serum were analyzed by ELISA ( n = 6). Data represent as mean ± SD. Compared to con, **** p < 0.0001; compared to a concentration of 0 μM, # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 and ns: not significant (one-way ANOVA with Dunnett’s post-hoc test)

Article Snippet: Anti-human IL-1β (p17, 12242S), anti-Phospho-IRF3 (Ser396) (4947), caspase-1 antibody (4199S) and anti-ASC antibody (sc-22, 514-R) were obtained from Cell Signaling Technology (Germany).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Concentration Assay