null2 cells  (InvivoGen)

Journal: PLOS One

Article Title: Ozone ultrafine bubble water sterilizes Porphyromonas gingivalis and neutralizes its virulence factors

doi: 10.1371/journal.pone.0334259

Figure Lengend Snippet: P. gingivalis LPS was added to distilled water (DW), air ultrafine bubble water (AUFBW), or 3.03 ppm ozone ultrafine bubble water (OUFBW). (A) HEK-Blue human Toll-like receptor 2 (hTLR2), (B) HEK-Blue hTLR4, and (C) HEK-Blue null2 cells were stimulated with the LPS. Pam3CSK4 (Pam), a TLR2 agonist, was used as the control. Secreted alkaline phosphatase (SEAP) levels were quantified using spectrophotometry at 620 nm. The data represent means ± SD of sextuplicate experiments and were evaluated by one-way analysis of variance with Tukey’s multiple comparisons test. †, significant difference compared to DW group at P < 0.05. *, significant difference between the indicated groups at P < 0.05.

Article Snippet: HEK-Blue cells expressing human TLR2 (HEK-hTLR2), HEK-hTLR4 and HEK-null2 cells (InvivoGen) were cultured at 37 °C in 5% CO 2 .

Techniques: Control, Spectrophotometry

Journal: Redox Biology

Article Title: 3- O -trans- p -coumaroyl esterification enhances the anti-inflammatory effects of tormentic acid by targeting NF-κB signaling

doi: 10.1016/j.redox.2025.103731

Figure Lengend Snippet: Effects of TA and trans -TACE on NF-κB activation and oxidative stress-related gene expression in cellular and C. elegans models . (A) NF-κB activation in TLR4-expressing HEK-Blue hTLR4 cells and non-TLR4-expressing HEK-Blue null2 cells following a 24-h treatment with TNFα (10 ng/ml), LPS (10 ng/ml) and TA or trans -TACE (5 μM), normalized to the stress control ( n = 9). (B) Schematic overview of key signaling pathways regulating oxidative stress responses and immune defense in C. elegans . The IIS pathway regulates DAF-16 activity, controlling antioxidant and stress response genes ( sod-3, gpx-5 ). The p38 MAPK pathway, activated via TIR-1, modulates SKN-1-dependent detoxification and immune defense genes ( gcs-1 , gst-4 ). Crosstalk between these pathways ensures coordinated stress adaptation. (C) Relative mRNA expression of daf-16 , sod-3 , gpx-5 , skn-1 , gcs-1 and gst-4 in C. elegans following a 24-h treatment with 50 μM TA or trans -TACE and stress induction by 20 mM PQ, normalized to PQ control ( n = 5 biological, 4 technical replicates). Data are represented as means ± SD of 2–3 independent experiments × p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: The medium was supplemented with 10 % heat-inactivated FBS, 100 U/ml-100 μg/ml P–S and 100 μg/ml NormocinTM (InvivoGen), as well as cell line-specific antibiotics 1X HEK-Blue Selection for the HEK-Blue hTLR4 cell line and 100 μg/ml Zeocin for the HEK-Blue null2 cell line (both from InvivoGen).

Techniques: Activation Assay, Gene Expression, Expressing, Control, Protein-Protein interactions, Activity Assay

Journal: Redox Biology

Article Title: 3- O -trans- p -coumaroyl esterification enhances the anti-inflammatory effects of tormentic acid by targeting NF-κB signaling

doi: 10.1016/j.redox.2025.103731

Figure Lengend Snippet: Comparative effects of trans/ cis -TACE and cis -TACE on key targets of oxidative and inflammatory stress in cellular and C. elegans models. (A) LC-MS chromatograms comparing the cis and trans isoform content of trans -TACE, trans/ cis -TACE and cis -TACE. (B) NF-κB activation in TLR4-expressing HEK-Blue hTLR4 cells and non-TLR4-expressing HEK-Blue null2 cells following a 24-h treatment with TNFα (10 ng/ml), LPS (10 ng/ml) and trans/ cis -TACE or cis -TACE (5 μM), normalized to the stress control ( n = 9–12). (C) Concentrations of TNFα, IL-8, CCL2 and CXCL5 in LPS (250 ng/ml)-stimulated THP-1 cells following a 4-h treatment with 10 μM trans/ cis -TACE or cis -TACE, quantified by multiplex bead-based cytokine immunoassay ( n = 4–6). (D) Intracellular ROS levels in Caco-2 cells after 20 min of treatment with 10 μM trans/ cis -TACE, cis -TACE or quercetin and subsequent stress induction by 300 μM AAPH, normalized to stressor treatment ( n = 12). (E) Relative mRNA expression of daf-16 , gst-4 , gpx-5 , skn-1 , gcs-1 and sod-3 in C. elegans following a 24-h treatment with 50 μM trans/ cis -TACE or cis -TACE and stress induction by 20 mM PQ, normalized to PQ control ( n = 6 biological, 4 technical replicates). Data are represented as means ± SD of 2–3 independent experiments × p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: The medium was supplemented with 10 % heat-inactivated FBS, 100 U/ml-100 μg/ml P–S and 100 μg/ml NormocinTM (InvivoGen), as well as cell line-specific antibiotics 1X HEK-Blue Selection for the HEK-Blue hTLR4 cell line and 100 μg/ml Zeocin for the HEK-Blue null2 cell line (both from InvivoGen).

Techniques: Liquid Chromatography with Mass Spectroscopy, Activation Assay, Expressing, Control, Multiplex Assay

Journal: Redox Biology

Article Title: 3- O -trans- p -coumaroyl esterification enhances the anti-inflammatory effects of tormentic acid by targeting NF-κB signaling

doi: 10.1016/j.redox.2025.103731

Figure Lengend Snippet: Effects of TA and trans -TACE on NF-κB activation and oxidative stress-related gene expression in cellular and C. elegans models . (A) NF-κB activation in TLR4-expressing HEK-Blue hTLR4 cells and non-TLR4-expressing HEK-Blue null2 cells following a 24-h treatment with TNFα (10 ng/ml), LPS (10 ng/ml) and TA or trans -TACE (5 μM), normalized to the stress control ( n = 9). (B) Schematic overview of key signaling pathways regulating oxidative stress responses and immune defense in C. elegans . The IIS pathway regulates DAF-16 activity, controlling antioxidant and stress response genes ( sod-3, gpx-5 ). The p38 MAPK pathway, activated via TIR-1, modulates SKN-1-dependent detoxification and immune defense genes ( gcs-1 , gst-4 ). Crosstalk between these pathways ensures coordinated stress adaptation. (C) Relative mRNA expression of daf-16 , sod-3 , gpx-5 , skn-1 , gcs-1 and gst-4 in C. elegans following a 24-h treatment with 50 μM TA or trans -TACE and stress induction by 20 mM PQ, normalized to PQ control ( n = 5 biological, 4 technical replicates). Data are represented as means ± SD of 2–3 independent experiments × p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: The HEK-Blue hTLR4 reporter cell line, which stably co-expresses an NF-κB-inducible secreted embryonic alkaline phosphatase (SEAP) reporter gene and the human toll like receptor (TLR) 4 gene, along with its corresponding parental cell line, HEK-Blue null2, which only expresses the SEAP reporter gene, were obtained from InvivoGen (San Diego, CA, USA).

Techniques: Activation Assay, Gene Expression, Expressing, Control, Protein-Protein interactions, Activity Assay

Journal: Redox Biology

Article Title: 3- O -trans- p -coumaroyl esterification enhances the anti-inflammatory effects of tormentic acid by targeting NF-κB signaling

doi: 10.1016/j.redox.2025.103731

Figure Lengend Snippet: Comparative effects of trans/ cis -TACE and cis -TACE on key targets of oxidative and inflammatory stress in cellular and C. elegans models. (A) LC-MS chromatograms comparing the cis and trans isoform content of trans -TACE, trans/ cis -TACE and cis -TACE. (B) NF-κB activation in TLR4-expressing HEK-Blue hTLR4 cells and non-TLR4-expressing HEK-Blue null2 cells following a 24-h treatment with TNFα (10 ng/ml), LPS (10 ng/ml) and trans/ cis -TACE or cis -TACE (5 μM), normalized to the stress control ( n = 9–12). (C) Concentrations of TNFα, IL-8, CCL2 and CXCL5 in LPS (250 ng/ml)-stimulated THP-1 cells following a 4-h treatment with 10 μM trans/ cis -TACE or cis -TACE, quantified by multiplex bead-based cytokine immunoassay ( n = 4–6). (D) Intracellular ROS levels in Caco-2 cells after 20 min of treatment with 10 μM trans/ cis -TACE, cis -TACE or quercetin and subsequent stress induction by 300 μM AAPH, normalized to stressor treatment ( n = 12). (E) Relative mRNA expression of daf-16 , gst-4 , gpx-5 , skn-1 , gcs-1 and sod-3 in C. elegans following a 24-h treatment with 50 μM trans/ cis -TACE or cis -TACE and stress induction by 20 mM PQ, normalized to PQ control ( n = 6 biological, 4 technical replicates). Data are represented as means ± SD of 2–3 independent experiments × p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: The HEK-Blue hTLR4 reporter cell line, which stably co-expresses an NF-κB-inducible secreted embryonic alkaline phosphatase (SEAP) reporter gene and the human toll like receptor (TLR) 4 gene, along with its corresponding parental cell line, HEK-Blue null2, which only expresses the SEAP reporter gene, were obtained from InvivoGen (San Diego, CA, USA).

Techniques: Liquid Chromatography with Mass Spectroscopy, Activation Assay, Expressing, Control, Multiplex Assay