hek blue il 1β cells  (InvivoGen)

Journal: mAbs

Article Title: A novel insulin-like growth factor II-based masking domain for conditional activation of therapeutic antibodies

doi: 10.1080/19420862.2026.2668187

Figure Lengend Snippet: Effects of masking on anti-IL-1β antibody activity. (A) ELISA binding assays showing IL-1β binding by the masked antibody IGF-II-MMP2/9-TAVO103A and the native TAVO103A. The absorbance at 450 nm (OD 450 nm) was plotted versus antibody concentration (ng/mL). The experiments were performed in duplicate with the data presented as mean values ± sd. (B) IL-1β-driven HEK-Blue reporter assays measuring IL-1β neutralization by IGF-II-MMP2/9-TAVO103A and TAVO103A. Percent IL-1β activity (normalized to the maximal response induced by 1 ng/mL IL-1β) was plotted versus antibody concentration (ng/mL). The experiments were performed in duplicate with the data presented as mean values ± sd.

Article Snippet: HEK-Blue IL-1β reporter assays were performed using HEK-Blue IL-1β cells (InvivoGen, hkb-il1bv2), which expressed IL-1 receptor complexes and the same NF-κB/AP-1-inducible SEAP reporter.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Binding Assay, Concentration Assay, Neutralization

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: RAW-DualTM cells (rawd-ismip), THP1-DualTM cells (thpd-nfis), HEK-Blue TM IL-1β cells (hkb-il1bv2; InvivoGen), THP1-Null2 Cells (thp-nullz) and THP1-KO-GSDMD cells (thp-kogsdmdz) were obtained from InvivoGen (San Diego, CA, USA).

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: RAW-DualTM cells (rawd-ismip), THP1-DualTM cells (thpd-nfis), HEK-Blue TM IL-1β cells (hkb-il1bv2; InvivoGen), THP1-Null2 Cells (thp-nullz) and THP1-KO-GSDMD cells (thp-kogsdmdz) were obtained from InvivoGen (San Diego, CA, USA).

Techniques: Comparison

Journal: bioRxiv

Article Title: TAK1 integrates the NLRP1 inflammasome into the innate immune response to double-stranded RNA

doi: 10.64898/2026.01.23.701331

Figure Lengend Snippet: ( A-B ) Reconstitution of the NLRP1 inflammasome by co-transfecting 293T cells with plasmids encoding NLRP1, ASC, CASP1, and pro-IL-1β. Cells were treated with VbP (10μM), mock transfected, or co-transfected with HMW poly(I:C) (1μg/mL) or plasmids encoding ZAKα or CVB3 3C protease. Levels of bioactive IL-1β, reported as optical density (OD), were quantified using a HEK-Blue IL-1β reporter assay (A, see Methods). Immunoblotting of indicated proteins from lysates harvested 40-42 hours post-transfection (B). ( C-E ) NLRP1 inflammasome reconstituted 293T cells were co-transfected with empty vector, pRIG-I, pMDA5, or pTLR3 and treated with VbP (10μM), mock transfected, or transfected with HMW poly(I:C) (1μg/mL). Quantification of supernatant IL-1β levels (D) and immunoblotting (E) were carried out as previously described. Bioactive IFN-β levels were quantified using a HEK-Blue IFN reporter assay (C). ( F ) Schematic depicting drug (B/B) inducible murine RIG-I CARDs fused to FKBP12 F36V dimerizing domains (3xFV-N-RIG-I CARD ). ( G ) NLRP1 inflammasome reconstituted 293T cells were co-transfected with either an empty vector or the 3xFV-N-RIG-I CARD and treated with VbP (10μM) or B/B (10nM). IL-1β was quantified as in (A). Results are representative of two (A-B, E,), three (G), or four (C-D) independent experiments. Data are presented as mean ± SD. p-values were determined by two-way ANOVA with Tukey’s test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Supernatant IL-1β and type I IFN from 293T cells was quantified using the HEK-Blue IL-1β and IFN-α/β reporter cells (Invivogen), respectively.

Techniques: Transfection, Reporter Assay, Western Blot, Plasmid Preparation

Journal: bioRxiv

Article Title: TAK1 integrates the NLRP1 inflammasome into the innate immune response to double-stranded RNA

doi: 10.64898/2026.01.23.701331

Figure Lengend Snippet: ( A-B ) WT and indicated single or double knockout (KO) N/TERT-2G cells were primed with 10ng/mL TNFα for 24 hours and then treated with VbP (10μM), mock transfected, or transfected with HMW poly(I:C) (1μg/mL). Supernatants were harvested 24 hours following the addition of activating stimuli and IL-1β (A) and IFN-β (B) were measured using an antibody-based immunoassay (see Methods). ( C ) Immunoblotting for indicated proteins from lysates harvested 6 hours following mock or poly(I:C) transfection of unprimed WT and indicated KO N/TERT-2G cells. ( D-F ) NLRP1 reconstituted 293T cells were either treated with VbP (10μM) or co-transfected with an empty vector, pMAVS, or pTRIF. IL-1β (D) and IFN-β (E) levels were quantified as described in . Immunoblotting for indicated proteins was carried out as described in (C) from lysates harvested 40-42 hours post-transfection (F). Results are representative of two (A-C) or three (D-F) independent experiments. Data are presented as mean ± SD. p-values were determined by one-way ANOVA with Dunnett’s test (A-B) or by two-way ANOVA with Tukey’s test (D-E). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Supernatant IL-1β and type I IFN from 293T cells was quantified using the HEK-Blue IL-1β and IFN-α/β reporter cells (Invivogen), respectively.

Techniques: Double Knockout, Transfection, Western Blot, Plasmid Preparation

Journal: bioRxiv

Article Title: TAK1 integrates the NLRP1 inflammasome into the innate immune response to double-stranded RNA

doi: 10.64898/2026.01.23.701331

Figure Lengend Snippet: ( A-C ) N/TERT-2G cells were primed with 10ng/mL TNFα for 24 hours. Cells were incubated with either Ruxolitnib (Rux) (1μM) or IFN blocking antibody (anti-IFN) (1:50) for 1 hour prior to treatment with either VbP (10μM), recombinant IFN-β (rIFN-β), or HMW poly(I:C) (1μg/mL) and supernatants and cell pellets were harvested 10 hours post treatment. Expression of MX1, IFIT1 , and ISG15 in poly(I:C)-treated N/TERT-2G cells was determined by quantitative RT-PCR (A). IFN-β (B) and IL-1β (C) were quantified by ELISA. ( D-E ) WT and indicated KO N/TERT-2G cells were primed with 10ng/mL TNFα for 24 hours and treated with VbP (10μM) or HMW poly(I:C) (1μg/mL) for 24 hours. Supernatant IL-1β (D) and IFN-β (E) were quantified using an antibody-based immunoassay. ( F ) Immunoblotting from lysates generated from unprimed WT and indicated KO N/TERT-2G cells treated with VbP (10μM) or HMW poly(I:C) (1μg/mL) for 6 hours. Results are representative of two (A-F) independent experiments. Data are presented as mean ± SD. p-values were determined by one-way ANOVA with Dunnett’s test (A, D, and E) or two-way ANOVA with Sidak’s test (B-C). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Supernatant IL-1β and type I IFN from 293T cells was quantified using the HEK-Blue IL-1β and IFN-α/β reporter cells (Invivogen), respectively.

Techniques: Incubation, Blocking Assay, Recombinant, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot, Generated

Journal: bioRxiv

Article Title: TAK1 integrates the NLRP1 inflammasome into the innate immune response to double-stranded RNA

doi: 10.64898/2026.01.23.701331

Figure Lengend Snippet: ( A ) IL-1β levels were quantified by ELISA from supernatants of WT and indicated KO N/TERT-2G cells treated with VbP (10μM), ANS (1μM), or HMW poly(I:C) (1μg/mL) for 24 hours. ( B ) Immunoblotting for indicated proteins from lysates generated from N/TERT-2G cells treated as described in (A) 6 hours post-treatment. ( C ) Schematic depicting dsRNA activation of TAK1 and ribotoxic stress activation of ZAKa upstream of the NLRP1 inflammasome. ( D ) Primary keratinocytes from three independent donors were treated with Doramapimod (p38 inhibitor; 10μM) or Takinib (TAK1 inhibitor; 10μM) for 2 hours prior to treatment with VbP (10μM), ANS (1μM), or HMW poly(I:C) (1μg/mL) for 24 hours. Supernatant IL-1β levels were quantified by an antibody-based immunoassay. ( E-F ) NLRP1 reconstituted 293T cells were either treated with VbP (10μM) or co-transfected with pZAKα, pTAK1 WT, or pTAK1 K63W. Quantification of supernatant IL-1β using HEK-Blue IL-1β reporter cells (D). Immunoblotting for indicated proteins from lysates harvested 40-42 hours post-transfection (E). Results are representative of two (A-B) or three (D-F) independent experiments. Data are presented as mean ± SD. p-values were determined by two-way ANOVA with Sidak’s test (A, E) or by one-way ANOVA with Dunnett’s test (D) using GraphPad PRISM 10. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Supernatant IL-1β and type I IFN from 293T cells was quantified using the HEK-Blue IL-1β and IFN-α/β reporter cells (Invivogen), respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Generated, Activation Assay, Transfection

Journal: bioRxiv

Article Title: TAK1 integrates the NLRP1 inflammasome into the innate immune response to double-stranded RNA

doi: 10.64898/2026.01.23.701331

Figure Lengend Snippet: ( A ) Schematic depicting phosphorylation motifs in NLRP1’s N-terminal disordered region. ( B-C ) 293T cells were co-transfected with plasmids encoding MDA5 and WT or NLRP1 phophomotif mutants and treated with VbP (10μM) or HMW poly(I:C) (1μg/mL) (B and C) or co-transfected with plasmids encoding MAVS, TRIF, ZAKα, or TAK1 (C). ( D ) 293T cells were co-transfected with plasmids encoding MDA5 and WT NLRP1, WT CARD8, CARD8 NLRP1-DR , or CARD8 NLRP1-DR2×3A and treated with either treated with VbP (10μM) or HMW poly(I:C) (1μg/mL) or co-transfected plasmids encoding ZAKα or TAK1. Supernatant IL-1β levels were measured using an IL-1β bioassay (B-D). Results are representative of two (B and D) or three (C) independent experiments. Data are presented as mean ± SD. p-values were determined by two-way ANOVA with Tukey’s test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Supernatant IL-1β and type I IFN from 293T cells was quantified using the HEK-Blue IL-1β and IFN-α/β reporter cells (Invivogen), respectively.

Techniques: Phospho-proteomics, Transfection, Bioassay

Journal: bioRxiv

Article Title: TAK1 integrates the NLRP1 inflammasome into the innate immune response to double-stranded RNA

doi: 10.64898/2026.01.23.701331

Figure Lengend Snippet: ( A ) WT and indicated KO N/TERT-2G cells were treated with either VbP (10μM) or infected with either SINV AR86 or GW (MOI=20) for 48 hours. IL-1β was measured using an antibody-based immunoassay ( B ) Primary keratinocytes were treated with Doramapimod (10μM) or Takinib (10μM) for 2 hours prior to infection with SINV AR86 (MOI=20) for 48 hours. Supernatant IL-1β was measured as in (A). ( C-E ) WT and indicated KO N/TERT-2G cells were treated with siRNAs against ADAR or a non-targeting control (NTC) for 24 hours and primed with 10ng/mL TNFα. Supernatants were harvested 72 hours following siRNA treatment and IL-1β (E) and IFN-β (F) levels were measured using an antibody-based immunoassay. Immunoblotting for indicated proteins was carried out as described in (G) from lysates harvested 72 hours post transfection (G). Results are representative of two (A-E) independent experiments. Data are presented as mean ± SD. p-values were determined by one-way ANOVA with Dunnett’s test (A-B) or by two-way ANOVA with Sidak’s test (C-D) using GraphPad PRISM 10. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Supernatant IL-1β and type I IFN from 293T cells was quantified using the HEK-Blue IL-1β and IFN-α/β reporter cells (Invivogen), respectively.

Techniques: Infection, Control, Western Blot, Transfection