Journal: Annals of clinical biochemistry

Article Title: 3-Hydroxyanthranilic acid, an L-tryptophan metabolite, induces apoptosis in monocyte-derived cells stimulated by interferon-gamma.

doi: 10.1258/0004563011900461

Figure Lengend Snippet: FIGURE 1. Effects of kynureninepathwaymetaboliteson apoptosisin humanmononuclearcell lines (THP-1and U937). Cells were treated with one of the L-tryptophan±kynurenine pathway metabolites (200mmol/L) for 48h then apoptotic cells were analysed by ¯uorescence-activated cell sorting (FACS) scan ¯ow cytometry with annexin V binding as described in the text. Each bar represents the mean and standard error of the mean of results from ®ve separate experiments. *P<0´01 compared with control. L-KYN=L-kynurenine; 3-HK=3-hydroxykynurenine; 3-HAA=3- hydroxyanthranilic acid; QUIN=quinolinic acid; KYNA=kynurenic acid; XA=xanthurenic acid; AA=anthranilic acid; PIC=picolinic acid, CBA=cinnabarinic acid.

Article Snippet: THP-1, U937 (acute monocyte leukaemia cell line), MRC9 (human lung cell line), H4 (human brain neuroglioma cell line), U373MG (astrocytoma) and Wil-NS (B lymphocyte) cells were obtained from ATCC (Rockville MD, USA).

Techniques: FACS, Cytometry, Binding Assay, Control

Journal: Annals of clinical biochemistry

Article Title: 3-Hydroxyanthranilic acid, an L-tryptophan metabolite, induces apoptosis in monocyte-derived cells stimulated by interferon-gamma.

doi: 10.1258/0004563011900461

Figure Lengend Snippet: FIGURE 3. Ferrous or manganese ion-enhanced apoptosis of THP-1 (a) and U937 (b) cells in the presence of 3- hydroxyanthranilic acid (3-HAA). Cells were treated with 3-HAA and/or ferrous (100mmol/L) or manganese ions (100mmol/L) for 48 h. Apoptotic cells were analysed by ¯uorescence-activatedcell sorting (FACS) scan with annexin V binding.

Article Snippet: THP-1, U937 (acute monocyte leukaemia cell line), MRC9 (human lung cell line), H4 (human brain neuroglioma cell line), U373MG (astrocytoma) and Wil-NS (B lymphocyte) cells were obtained from ATCC (Rockville MD, USA).

Techniques: Binding Assay

Journal: Annals of clinical biochemistry

Article Title: 3-Hydroxyanthranilic acid, an L-tryptophan metabolite, induces apoptosis in monocyte-derived cells stimulated by interferon-gamma.

doi: 10.1258/0004563011900461

Figure Lengend Snippet: FIGURE 5. Effects of ferrous or manganese ions on apoptosis in interferon-g (IFN-g)-treated human monocyte- derived cells. Human THP-1 and U937 cells were treated with IFN-g and/or ferrous or manganese ions for 48 h. Apoptotic cells were analysed by ¯uorescence-activated cell sorting (FACS) scan with annexin V binding. *P<0´05 and **P<0´01 compared with control. +P<0´01 compared with ferrous or manganese ions plus IFN-g treatment. CHX=cycloheximide

Article Snippet: THP-1, U937 (acute monocyte leukaemia cell line), MRC9 (human lung cell line), H4 (human brain neuroglioma cell line), U373MG (astrocytoma) and Wil-NS (B lymphocyte) cells were obtained from ATCC (Rockville MD, USA).

Techniques: Derivative Assay, FACS, Binding Assay, Control

Journal: Annals of clinical biochemistry

Article Title: 3-Hydroxyanthranilic acid, an L-tryptophan metabolite, induces apoptosis in monocyte-derived cells stimulated by interferon-gamma.

doi: 10.1258/0004563011900461

Figure Lengend Snippet: FIGURE 6. Schematic overview of kynurenine pathway metabolism when THP-1 and U937 cells were treated with interferon-g (IFN-g). L-Trp=L-tryptophan; IDO=indoleamine-2,3-dioxygenase; L-KYN=L-kynurenine; 3-HK=3- hydroxykynurenine; 3-HAA=3-hydroxyanthranilic acid; QUIN=quinolinic acid; CBA=cinnabarinic acid.

Article Snippet: THP-1, U937 (acute monocyte leukaemia cell line), MRC9 (human lung cell line), H4 (human brain neuroglioma cell line), U373MG (astrocytoma) and Wil-NS (B lymphocyte) cells were obtained from ATCC (Rockville MD, USA).

Techniques:

Journal: Nature Communications

Article Title: A Pseudomonas aeruginosa quorum-sensing metabolite manipulates macrophage ferroptosis through a methylation pathway

doi: 10.1038/s41467-025-65142-y

Figure Lengend Snippet: A Growth curves of PAO1, Δ pqsR , and Δ pqsR + strains. B Lactate dehydrogenase (LDH) assay for cell death in RAW264.7 macrophages infected with PAO1, Δ pqsR , and Δ pqsR + (Δ pqsR mutant expressing pqsR from a rescue construct pMiniCTX1- pqsR ) Bac (bacteria cultures) or Sup (cell-free supernatants). Multiplicity of infection (MOI) = 100. Data represent the means ± SD ( n = 3, two-way ANOVA with Tukey’s multiple comparisons test). C ELISA quantification of PQS concentrations measured in PAO1, Δ pqsR , and Δ pqsR + supernatants. Data represent the means ± SD ( n = 3, one-way ANOVA with Tukey’s multiple comparisons test). D LDH assay for cell death in RAW264.7 macrophages treated with different concentrations of PQS for 24 h, untreated macrophages (Ctrl), and macrophages treated with DMSO as the negative control. Data represent the means ± SD ( n = 3, one-way ANOVA with Tukey’s multiple comparisons test). E CCK-8 assay for cell viability in RAW264.7 macrophages treated with different concentrations of PQS for 24 h; IC 50 (PQS) = 14.98 μg/mL. Data represent the means ± SD ( n = 3). F Microscopic observation of the effect of concentration of IC 50 (PQS) exposure for 24 h on RAW264.7 macrophage morphology. Scale bar = 100 µm. G Transcription electron microscopy (TEM) observation of the effect of 10 μg/mL PQS exposure on RAW264.7 macrophage morphology. Nu: cell nucleus, Au-L: autolysosome. 15/40KK: × 15000/ 40,000 magnifications. Scale bar = 2/0.5 μm. H CCK-8 assay for cell viability in BMDM, THP-1, U937, A549, and BEAS-2B cells treated with 10 μg/mL PQS for 24 h. Data represent the means ± SD ( n = 3). Source data are provided as a Source Data file.

Article Snippet: The RAW264.7 murine macrophages (ATCC ® TIB-71 TM ), THP-1 human monocyte (ATCC ® TIB-202 TM ), U937 human leukaemia (ATCC ® CRL-3253 TM ), A549 human epithelium (ATCC®CCL-185TM), and BEAS-2B human bronchial epithelium (ATCC ® CRL-3588 TM ) cell lines were grown in Dulbecco’s Modified Eagle (DMEM) or Roswell Park Memorial Institute (RPMI)-1640 medium supplemented with 10% foetal bovine serum (FBS) and 1% penicillin/streptomycin (PS) (Gibco, Texas, USA), followed by incubation at 37 °C in a 5% CO 2 incubator.

Techniques: Lactate Dehydrogenase Assay, Infection, Mutagenesis, Expressing, Construct, Bacteria, Enzyme-linked Immunosorbent Assay, Negative Control, CCK-8 Assay, Concentration Assay, Electron Microscopy

Journal: bioRxiv

Article Title: Macrophage-based assays for the in vitro testing of the anti-inflammatory activity of mesenchymal stem cell-based products

doi: 10.64898/2026.03.04.709479

Figure Lengend Snippet: The anti-inflammatory effect of ASC secretome seeded in transwells (tw) in various LPS-stimulated macrophage models. A-C: the mRNA expression of inflammatory markers in the U937 macrophages. D-F: the mRNA expression of inflammatory markers in the THP-1 macrophages. G-I: the mRNA expression of inflammatory markers in the RAW264.7 macrophages. A-F: The data are presented as the mean relative mRNA expression ± standard deviation from four independent experiments. G-I: The data are presented as the mean relative mRNA expression ± standard deviation from seven independent experiments. The statistical analysis was performed applying the one-sample t-test (comparison with the reference value of 100%). The statistical significance of MΦ versus MΦ + LPS is displayed as “#” and the statistical significance of MΦ + LPS versus MΦ + LPS + ASC is displayed as “*”. The significance is indicated as *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001; ns, not significant (# is used analogically).

Article Snippet: The U937 (human acute monocytic leukaemia cell line, Merck, cat. # 85011440-1VL) and THP-1 (human acute monocytic leukaemia cell line, Merck, cat. # 88081201-1VL) cells were cultured in RPMI 1640 medium (Biosera) supplemented with 10% FBS, 2 mM L-glutamine, 1% penicillin/streptomycin and 1 mM sodium pyruvate (Merck); the THP-1 medium further contained 4.5 g/l D-glucose (Thermo Fisher) and 0.05 mM β-mercaptoethanol (Merck).

Techniques: Expressing, Standard Deviation, Comparison

Journal: bioRxiv

Article Title: Macrophage-based assays for the in vitro testing of the anti-inflammatory activity of mesenchymal stem cell-based products

doi: 10.64898/2026.03.04.709479

Figure Lengend Snippet: the anti-inflammatory effect of the ASC secretome on the LPS-pre-activated U937 macrophages. The U937 cells were briefly (3 h) pre-activated with LPS prior to the seeding of the ASC. A-C: The effect of the ASC secretome on the mRNA expression of the M1 markers. D-H: The effect of the ASC secretome on the mRNA expression of the M2 markers. The data are presented as the mean relative mRNA expression ± the standard deviation from three independent experiments. The statistical analysis was performed applying the one-sample t-test (comparison with the reference value of 100%). The statistical significance of MΦ versus MΦ + IL-4 + IL-13 is displayed as “#” and the statistical significance of MΦ + IL-4 + IL-13 versus MΦ + IL-4 + IL-13 + ASC is displayed as “*”. The significance is indicated as *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001; ns, not significant (# is used analogically).

Article Snippet: The U937 (human acute monocytic leukaemia cell line, Merck, cat. # 85011440-1VL) and THP-1 (human acute monocytic leukaemia cell line, Merck, cat. # 88081201-1VL) cells were cultured in RPMI 1640 medium (Biosera) supplemented with 10% FBS, 2 mM L-glutamine, 1% penicillin/streptomycin and 1 mM sodium pyruvate (Merck); the THP-1 medium further contained 4.5 g/l D-glucose (Thermo Fisher) and 0.05 mM β-mercaptoethanol (Merck).

Techniques: Expressing, Standard Deviation, Comparison

Journal: bioRxiv

Article Title: Macrophage-based assays for the in vitro testing of the anti-inflammatory activity of mesenchymal stem cell-based products

doi: 10.64898/2026.03.04.709479

Figure Lengend Snippet: The effect of the ASC secretome on the M2 polarisation of U937. A-E: the mRNA expression of the various M2 markers. The data are presented as the mean relative mRNA expression ± the standard deviation from four independent experiments. The statistical analysis was performed applying the one-sample t-test (comparison with the reference value of 100%). The statistical significance of MΦ versus MΦ + IL-4 + IL-13 is displayed as “#” and the statistical significance of MΦ + IL-4 + IL-13 versus MΦ + IL-4 + IL-13 + ASC is displayed as “*”. The significance is indicated as *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001; ns, not significant (# is used analogically).

Article Snippet: The U937 (human acute monocytic leukaemia cell line, Merck, cat. # 85011440-1VL) and THP-1 (human acute monocytic leukaemia cell line, Merck, cat. # 88081201-1VL) cells were cultured in RPMI 1640 medium (Biosera) supplemented with 10% FBS, 2 mM L-glutamine, 1% penicillin/streptomycin and 1 mM sodium pyruvate (Merck); the THP-1 medium further contained 4.5 g/l D-glucose (Thermo Fisher) and 0.05 mM β-mercaptoethanol (Merck).

Techniques: Expressing, Standard Deviation, Comparison

Journal: bioRxiv

Article Title: Macrophage-based assays for the in vitro testing of the anti-inflammatory activity of mesenchymal stem cell-based products

doi: 10.64898/2026.03.04.709479

Figure Lengend Snippet: The effect of the ASC secretome on the mRNA expression of the M1 markers in the U937 M2 macrophage model. The data are presented as the mean relative mRNA expression ± the standard deviation from four independent experiments. The statistical analysis was performed applying the one-sample t-test (comparison with the reference value of 100%). The statistical significance of MΦ versus MΦ + IL-4 + IL-13 is displayed as “#” and the statistical significance of MΦ + IL-4 + IL-13 versus MΦ + IL-4 + IL-13 + ASC is displayed as “*”. The significance is indicated as *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001; ns, not significant (# is used analogically).

Article Snippet: The U937 (human acute monocytic leukaemia cell line, Merck, cat. # 85011440-1VL) and THP-1 (human acute monocytic leukaemia cell line, Merck, cat. # 88081201-1VL) cells were cultured in RPMI 1640 medium (Biosera) supplemented with 10% FBS, 2 mM L-glutamine, 1% penicillin/streptomycin and 1 mM sodium pyruvate (Merck); the THP-1 medium further contained 4.5 g/l D-glucose (Thermo Fisher) and 0.05 mM β-mercaptoethanol (Merck).

Techniques: Expressing, Standard Deviation, Comparison