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, representation of the proteins with denaturing curves significantly altered (red) or unaltered (black) by 3,2-HPP treatment of THP1-Dual™ cells, using thermal proteome profiling (two combined experiments, proteomic coverage: 4301, NPARC test); b, protein-protein interaction network of proteins with denaturation curves altered by 3,2-HPP treatment (red) and the transcriptional regulators of 3, 2-HPP impacted genes in M3-9-M tumours (yellow); c, impact of 3,2-HPP treatment of THP1-Dual™ cells on the denaturation curve of GSDMD (two-combined experiments, NPARC test); d, NF-κB induction in THP1-Dual™ reporter cells treated with conditioned media harvested from WT or Gsdmd- KO THP1 cells treated with LPS ± HPP metabolites ± IL-1α and −1β neutralizing antibody (three combined experiments, one-way ANOVA with Bonferroni multiple comparison test); e-h, [secreted IL-1α and −1β] in conditioned media harvested from THP-1 WT or Gsdmd -KO cells, or human PBMCs, cultured for 16 hours in LPS ± HPPs (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); i-l, [secreted IL-1α and −1β] in conditioned media harvested from THP-1 WT vs. Gsdmd -KO cells, or human PBMCs, cultured for 16 hours in LPS and NG ± HPPs (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); m, western blot of THP-1 cells treated with LPS ± NG ± HPPs (representative of two experiments).

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, Cell Culture, Western Blot

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 Immunology

Article Title: Macrophage and IL-6 signaling modulate multiple myeloma progression and response to metformin and CAR-T cell therapy

doi: 10.3389/fimmu.2026.1760136

Figure Lengend Snippet: Effects of IL-6 on cellular proliferation. (A) RNA and protein expression of IL-6 receptor in various cancer cells from the HPA database ( https://www.proteinatlas.org/ENSG00000160712-IL6R/cell+line , accessed on 29 Nov 2025). (B) Luciferase reporter assay for measuring RPMI 8226 cells activity with or without THP-1 cells, and IL-6 (5 ng/ml) for 48 hours. The seeding number of MM cells was 10,000. THP-1 cell numbers (THP-1 N.) were varied at 2000 (2K), 4000 (4K), 8000 (8K), and 10,000 (10K). (C) THP-1 cells were stimulated with IL-6 at concentrations of 0, 5, or 10 ng/ml for 24 hours. At the end of incubation, mRNA expression levels of iNOS and CD163 in THP-1 cells were quantified by qRT-PCR, and the ratio was determined by comparing the 2-ΔΔCt values of CD163 and iNOS. N = 9. ****p <0.0001.

Article Snippet: Anti-human IL-6 was purchased from Invivogen (Cat #mabg-hil6-3).

Techniques: Expressing, Luciferase, Reporter Assay, Activity Assay, Incubation, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Macrophage and IL-6 signaling modulate multiple myeloma progression and response to metformin and CAR-T cell therapy

doi: 10.3389/fimmu.2026.1760136

Figure Lengend Snippet: Effects of metformin on RPMI 8226 cells with macrophages. (A) Luciferase reporter assay of RPMI 8226 cells following PBS control (Ctrl), metformin (Met. 1mM) or IL-6 (5ng/ml) treatment for 72 hours. (B) QRT-PCR analysis of CD163 and iNOS expression in THP-1 cells under various treatment conditions for 72 hours. (C) Flow cytometry analysis of CD126 expression on RPMI 8226 cells following 72 hours of metformin treatment with or without macrophages. (D) Luciferase reporter assay of RPMI 8226 cells under various treatment conditions at 72 hours. N = 4-9. *p<0.05, **p <0.01, ***p <0.001, ****p <0.0001.

Article Snippet: Anti-human IL-6 was purchased from Invivogen (Cat #mabg-hil6-3).

Techniques: Luciferase, Reporter Assay, Control, Quantitative RT-PCR, Expressing, Flow Cytometry