thp1 dualtm cells thpd nfis  (InvivoGen)

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, immune cell types in the TME identified through scRNAseq of CD45 + cells isolated from orthotopically implanted M3-9-M OVA cells; b, number of immune cell types detected in M3-9-M OVA tumours through scRNAseq experiment (n = 2 pooled samples per experimental groups); c, number of significantly impacted genes obtained from pairwise comparisons (Wilcoxon rank sum test); d, circos plot of all significantly impacted genes, obtained from pairwise comparisons of all cell types measured, that overlap with each other through identical or shared pathways; e, bar graph of transcriptional regulators of 3,2-HPP impacted genes; f-m, NF-κB and IRF induction in mouse RAW-Dual™ cells and human THP1-Dual™ cells with different pathogen recognition receptor agonists in the presence of HPP metabolites (HI = heat inactivated, three combined experiments, error bars represent standard error, one-way ANOVA 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: Isolation

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, NF-κB activity in THP1-Dual™ cells treated with LPS in the presence of vehicle or HPP metabolites for 0-24 hours (two combined experiments, error bars represent standard error); b-e, NF-κB or IRF activity in THP1-Dual™ cells treated with different inducers in the presence or absence of HPP isomers for 16 hours (three combined experiments, error bars represent standard error, one-way ANOVA 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, 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: 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, 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: 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: Stress-responsive Mycobacterium tuberculosis subpopulations manipulate macrophage polarization and can be targeted to limit inflammation

doi: 10.64898/2026.03.13.711673

Figure Lengend Snippet: GroEL2 mRuby phenotypic variation predicts different macrophage responses (A–D) Snapshot images of THP-1 cells infected with GroEL2 mRuby -GFP cyt reporter at 24 and 48 hpi (n = 3). GroEL2 mRuby -Dim (A, C) and -Bright (B, D) intracellular bacilli are shown (dashed boxes). Maximum intensity projections of four images from a z-stack of 8 µm; bright-field (blue), GFP cyt (green), and GroEL2 mRuby (magenta) channels are merged. Scale bars, 10 µM. (E) GroEL2 mRuby normalized to GFP cyt mean fluorescence of exponential-phase (EP) bacilli and intracellular infectious foci at 24 and 48 hpi. Lines represent mean ± SD (n = 2, with cell number shown at the bottom of the graphs). Significance by nested one-way ANOVA (P = 0.034) followed by Tukey multiple comparisons test (adjusted P values). (F) Time course of GroEL2 mRuby /GFP cyt ratio for single infectious foci, classified as Dim (mean ratio < 1, n = 31) or Bright (mean ratio > 1, n = 20). Dashed line indicates the threshold ratio. Data are from three independent experiments. (G) Pearson’s correlation between intracellular growth rate and GroEL2 mRuby /GFP cyt ratio of individual intracellular infectious foci over the infected cell lifetime (n = 64 from two independent experiments). The dotted line indicates the threshold ratio. Simple linear regression fit (solid line, R 2 = 0.36, P < 0.0001) and 95% confidence bands (dashed lines). (H) Flow cytometry analysis of infected THP-1 macrophages at 24 hpi, sorted into GroEL2 mRuby -Dim (gray) and -Bright (pink) subpopulations based on intracellular fluorescence. Representative overlays are normalized to 10,000 events (n = 7 independent experiments). (I) Bacterial load in GroEL2 mRuby -Dim and -Bright THP-1 subpopulations at 24 hpi. Lines represent mean ± SD (n = 7). Significance by two-tailed unpaired t -test (95% confidence level). (J) Immunoluminometric quantification of cytokines at 48 hpi in GroEL2 mRuby -Dim (gray) and -Bright (pink) THP-1 subpopulations sorted at 24 hpi as in (n = 7). Significance by multiple paired t -tests with Holm-Sidak correction (adjusted P values). (K) Quantification of oxidative burst in THP-1 in subpopulations sorted at 48 hpi based on intracellular GroEL2 mRuby levels. Lines represent mean ± SD (n = 4 independent experiments). Significance by two-tailed unpaired t -test (95% confidence level). (L) Representative snapshot images of THP-1 cells infected with GroEL2 mRuby -GFP cyt reporter at 48 hpi, stained with annexin V (A5) and DRAQ7 (D7) (n = 3 independent experiments). Maximum intensity projections of four images from a z-stack of 6 µm; bright-field (blue), A5 (cyan), GFP cyt (green), GroEL2 mRuby and D7 (magenta) channels are merged. Scale bars, 10 µm. (M) GroEL2 mRuby /GFP cyt ratio for single intracellular infectious foci at 48 hpi in THP-1 macrophages, classified according to A5 and D7 negative (–) or positive (+) staining. Lines represent mean ± SD (n = 3, with cell number shown at the bottom of the graphs). Significance by two-way ANOVA (P = 0.009), followed by multiple comparisons with two-stage linear step-up procedure of Benjamini, Krieger and Yekutieli (individual P values).

Article Snippet: THP-1 DualTM and THP-1 DualTM MyD88 knock-out cells (InvivoGen) were grown in RPMI 1640 (Gibco) supplemented with 10% FBS as above, 25 mM HEPES, 100 μg/mL Pen-Strep, 100 μg/mL Normocin, 10 μg/mL Blasticidin, and 100 μg/mL Zeocin.

Techniques: Infection, Fluorescence, Flow Cytometry, Two Tailed Test, Staining

Journal: bioRxiv

Article Title: Stress-responsive Mycobacterium tuberculosis subpopulations manipulate macrophage polarization and can be targeted to limit inflammation

doi: 10.64898/2026.03.13.711673

Figure Lengend Snippet: GroEL2 H triggers NF-κB and IRF signaling and can be targeted to decrease inflammation (A) Immunoblotting of M. tuberculosis extracellular (EC) and intracellular (IC) fractions using anti-HSP65 mouse monoclonal IgG 2a . Control (CT) and rv2224c -silenced (si) strains in the absence (–) or presence (+) of ATC inducer (200 ng/mL). Loaded sample amounts in percentage, high (H) and low (L) molecular-weight GroEL2 bands, and Ag85 as a control are indicated. (B and C) Activation of NF-κB (B) and IRF (C) in WT (circles) and KO MyD88 (squares) dual-reporter THP-1 cells 24 hpi. Cells were infected with either M. tuberculosis CT (filled symbols) or rv2224c si (open symbols) at MOI 2 for 2 hours, 4 days after induction of silencing. Absence (–) or presence (+) of inducer and TLR4 inhibitor (760 nM) is indicated. NF-κB_SEAP (B) and ISRE_Lucia (C) signals are normalized to uninfected cells. Significance by one-way ANOVA (P < 0.0001), followed by Šidák’s multiple comparisons test (adjusted P values). Lines represent mean ± SEM (n = 3). (D–F) Activation of NF-κB (D) and IRF (E), and TNF-α production (F) in WT THP-1 dual-reporter cells infected with M. tuberculosis H37Rv (MOI 5 or 10) for 3 hours. Cells were left untreated (circles) or treated with mr38548 (MIC: 0.88 µg/mL; inverted triangles) or RIF (MIC: 0.076 µg/mL; triangles) at different concentrations relative to the MIC (x) for 24 hours. Values are normalized to uninfected and untreated cells. Significance by non-parametric Friedman test: P = 0.016 (D) and P < 0.0001 (E and F), followed by Dunn’s multiple comparisons test (adjusted P values). Lines represent mean ± SEM (n ≥ 6). (G–I) Activation of NF-κB (G) and IRF (H), and TNF-α production (I) in WT THP1 dual-reporter cells infected with M. tuberculosis H37Rv (MOI 5 or 10; circles) or stimulated with LPS (100 ng/mL; diamonds) for 5 hours. Cells were left untreated or exposed to different concentrations of monoclonal anti-HSP65 IgG 2a . Values are normalized to uninfected and untreated cells. Significance by mixed-model one-way ANOVA: P = 0.0014 (G), P = 0.0002 (H), and P = 0.0274 (I), followed by Šidák’s multiple comparisons test (adjusted P values). Lines represent mean ± SEM (n ≥ 3).

Article Snippet: THP-1 DualTM and THP-1 DualTM MyD88 knock-out cells (InvivoGen) were grown in RPMI 1640 (Gibco) supplemented with 10% FBS as above, 25 mM HEPES, 100 μg/mL Pen-Strep, 100 μg/mL Normocin, 10 μg/mL Blasticidin, and 100 μg/mL Zeocin.

Techniques: Western Blot, Control, Molecular Weight, Activation Assay, Infection

Journal: Frontiers in Microbiology

Article Title: Modulation of LPS-associated virulence activity for reduction of periodontal inflammatory burden

doi: 10.3389/fmicb.2026.1728315

Figure Lengend Snippet: Modulation of NF-κB and IRF signaling pathways in THP-1 Dual cells by LL-37 and Polymyxin B. (A) NF-κB activity quantified using the Quanti-Blue assay. (B) IRF activity measured using the Quanti-Luc assay. Data represent mean ± SD. Statistical significance relative to the untreated condition was determined using a paired t -test with Benjamini-Hochberg correction for multiple comparisons (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: THP-1 monocytes and THP-1 DualTM reporter cells (InvivoGen, US) were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum and 1% penicillin–streptomycin and then equilibrated in antibiotic-free medium before stimulation.

Techniques: Protein-Protein interactions, Activity Assay

Journal: Frontiers in Microbiology

Article Title: Modulation of LPS-associated virulence activity for reduction of periodontal inflammatory burden

doi: 10.3389/fmicb.2026.1728315

Figure Lengend Snippet: Cytokine secretion profiles in THP-1 cells stimulated with pooled clinical samples and purified LPS controls, with and without modulation by LL-37 and polymyxin B. Data are presented as mean ± SD. Statistical significance relative to the corresponding untreated condition was determined using a paired t -test adjusted for multiple comparisons via the Benjamini-Hochberg method (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: THP-1 monocytes and THP-1 DualTM reporter cells (InvivoGen, US) were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum and 1% penicillin–streptomycin and then equilibrated in antibiotic-free medium before stimulation.

Techniques: Purification