Journal: Food & Nutrition Research

Article Title: Urolithin A supplementation alleviates osteogenic disfunction and promotes bone fracture healing in inflammatory environments

doi: 10.29219/fnr.v70.13033

Figure Lengend Snippet: Sequences of primers for real-time quantitative PCR analysis

Article Snippet: The antibody against IL-1 β (#HY- P80720 ) was purchased from MedChemExpress.

Techniques: Real-time Polymerase Chain Reaction

Journal: Food & Nutrition Research

Article Title: Urolithin A supplementation alleviates osteogenic disfunction and promotes bone fracture healing in inflammatory environments

doi: 10.29219/fnr.v70.13033

Figure Lengend Snippet: UA alleviated pro-inflammation level and the M1 macrophage activation under TNF- α induced inflammation microenvironment. (a) CCK-8 assay was utilised to assess the cell viability of RAW264.7 of UA on 12, 24 and 48 h. (b, c) The concentration of IL-1 β and IL-6 in the medium of RAW264.7 after 12 h TNF- α stimulation with or without UA by ELISA. (d) The proteins of RAW264.7 were measured by western blot after 1 day and quantitative analysis. (e) The genes were measured by qRT-PCR after 1 day. (f) Immunofluorescence staining for iNOS after 1 day after TNF- α stimulation with or without UA. Scale bars, 100 μm. (g) The quantitative analysis of Immunofluorescence by image J. All of the experiments were independently accomplished by three times. * P < 0.05, ** P < 0.01 compared to the control group.

Article Snippet: The antibody against IL-1 β (#HY- P80720 ) was purchased from MedChemExpress.

Techniques: Activation Assay, CCK-8 Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining, Control

Journal: Food & Nutrition Research

Article Title: Urolithin A supplementation alleviates osteogenic disfunction and promotes bone fracture healing in inflammatory environments

doi: 10.29219/fnr.v70.13033

Figure Lengend Snippet: UA accelerated TNF- α induced impaired bone fracture healing in a mice femur fracture model. (a) Experiment flow about animal study. (b) The representative μCT images of mouse femurs from the femur fracture + PBS, femur fracture + TNF- α and femur fracture + TNF- α + UA (50 mpk) group. (c) Graphic illustrations of BV/TV, Tb.N, Tb.Th and Tb.Sp in the indicated groups. (d) Histological section from femur fracture region stained with H&E and Masson staining. Scale bar, 500 μm. (e) Immunohistochemistry of COL1A1 and IL-1 β of femur fracture region. Scale bar, 500 μm. (f, g) The quantitative results of immunohistochemistry by image J. (h) Histological scores of three groups. All of the experiments were independently accomplished by five times. All error bars represent SDs. * P < 0.05, ** P < 0.01 compared to the control group.

Article Snippet: The antibody against IL-1 β (#HY- P80720 ) was purchased from MedChemExpress.

Techniques: Staining, Immunohistochemistry, Control

Journal: Scientific Reports

Article Title: A rabbit model of clinically relevant mucosal injury induced by nasogastric tube intubation

doi: 10.1038/s41598-026-36598-9

Figure Lengend Snippet: The process of NGT intubation in the New Zealand rabbit . ( a ) Measuring intubation depth, ( b ) The supine position for intubation, ( c ) Inserting NGT, ( d ) Fixing the NGT on the nose of the rabbit via surgical suture, ( e ) The fixed NGT, ( f ) Wearing an Elizabeth collar.

Article Snippet: The sections of nasal septum mucosa were deparaffinized, dehydrated and stained with primary antibodies against IL-1β (1:500, Bioss, bs-0812R, China), IL-8 (1:500, Bioss, bs-0780R, China), and IL-6 (1:100, Servicebio, GB11117, China).

Techniques:

Journal: Scientific Reports

Article Title: A rabbit model of clinically relevant mucosal injury induced by nasogastric tube intubation

doi: 10.1038/s41598-026-36598-9

Figure Lengend Snippet: The expression of inflammatory cytokines. ( a ) Immunofluorescence staining images of IL-1β, IL-8 and IL-6 in the nasal septum mucosa, and the corresponding relative fluorescence intensity of ( b ) IL-1β ( c ) IL-8 and ( d ) IL-6. The relative mRNA expression of inflammatory cytokines including ( e ) IL-1β ( f ) IL-8 and ( g ) IL-6 in the nasal septum mucosa after extubation. Error bars represent ± SD ( n = 4), * p <0.05, ** p <0.01 and *** p <0.001.

Article Snippet: The sections of nasal septum mucosa were deparaffinized, dehydrated and stained with primary antibodies against IL-1β (1:500, Bioss, bs-0812R, China), IL-8 (1:500, Bioss, bs-0780R, China), and IL-6 (1:100, Servicebio, GB11117, China).

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence

Journal: Frontiers in Immunology

Article Title: Artemisinin exerts anti-inflammatory effects in osteoarthritis through the inhibition of TGF-β1 signaling

doi: 10.3389/fimmu.2026.1717045

Figure Lengend Snippet: Workflow of the mendelian randomization study. The diagram illustrates the study design and analysis process, including the selection of genetic instruments for inflammatory cytokines, coordination with Osteoarthritis (OA) genome-wide association study (GWAS) data, primary inverse variance weighting (IVW) analysis, sensitivity analysis, evaluation of heterogeneity and pleiotropy, and robustness assessment through leave-one-out analysis.

Article Snippet: The membranes were blocked and subsequently incubated overnight with primary antibodies against IL-1β and MMP-13 (both from Proteintech, China), followed by incubation with an HRP-conjugated goat anti-rabbit secondary antibody (Proteintech, China).

Techniques: Selection, GWAS

Journal: Frontiers in Immunology

Article Title: Artemisinin exerts anti-inflammatory effects in osteoarthritis through the inhibition of TGF-β1 signaling

doi: 10.3389/fimmu.2026.1717045

Figure Lengend Snippet: Effects of ART and TGF-β1 on the expression of IL-1β and MMP-13. (A) Relative mRNA expression of IL1B in each group, analyzed by RT-qPCR. (B) Relative mRNA expression of MMP13 in each group, analyzed by RT-qPCR. (C) Western blot analysis of IL-1β and MMP-13 protein expression. (D) Quantification of protein band intensity by densitometric analysis (normalized to β-actin). (E) Immunofluorescence staining showing the expression and localization of IL-1β and MMP-13 in cells (green indicates target protein, blue indicates DAPI nuclear staining). Scale bar = 100 μm. (F) Quantitative analysis of mean fluorescence intensity in immunofluorescence images. Data are presented as mean ± SD from three independent biological replicates (n=3). Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s multiple-comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The membranes were blocked and subsequently incubated overnight with primary antibodies against IL-1β and MMP-13 (both from Proteintech, China), followed by incubation with an HRP-conjugated goat anti-rabbit secondary antibody (Proteintech, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, Fluorescence

Journal: Frontiers in Immunology

Article Title: Artemisinin exerts anti-inflammatory effects in osteoarthritis through the inhibition of TGF-β1 signaling

doi: 10.3389/fimmu.2026.1717045

Figure Lengend Snippet: Artemisinin suppresses the pro-inflammatory effects of TGF-β1 in OA chondrocytes. (A) Relative mRNA expression of IL1B in each group, analyzed by RT-qPCR. (B) Relative mRNA expression of MMP13 in each group, analyzed by RT-qPCR. (C) Western blot analysis of IL-1β and MMP-13 protein expression. (D) Quantification of protein band intensity by densitometric analysis (normalized to β-actin). (E) Immunofluorescence staining showing the expression and localization of IL-1β and MMP-13 (green indicates target protein; blue indicates DAPI nuclear staining). Scale bar = 100 μm. (F) Quantitative analysis of mean fluorescence intensity in immunofluorescence images. Data are presented as mean ± SD from three independent biological replicates (n=3). Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s multiple-comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The membranes were blocked and subsequently incubated overnight with primary antibodies against IL-1β and MMP-13 (both from Proteintech, China), followed by incubation with an HRP-conjugated goat anti-rabbit secondary antibody (Proteintech, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, Fluorescence

Journal: Frontiers in Immunology

Article Title: TGFβ1 attenuates microglial IL1β release through inhibition of NLRP3 inflammasome priming

doi: 10.3389/fimmu.2025.1623643

Figure Lengend Snippet: TGFβ1 significantly reduces LPS-induced increases in NLRP3, CASP1, and IL1β protein levels in BV2 cells. After treatment with TGFβ1 (5 ng/ml), LPS (1 µg/ml) or the combination of both factors for 6 h and 12 h, protein levels were analyzed using western blotting. Representative western blot images for both experimental time points are depicted for NLRP3 (A) , CASP1 (C) , and IL1 β (E) . Quantifications after normalization using β-Actin are presented for NLRP3 (B) , CASP1 (D) , and IL1β (F) . Data are given as means ± SEM from at least three independent experiments. P-values derived from one-way ANOVA followed by Tukey´s multiple comparison tests are * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: They were later incubated with primary antibodies against Il1β (AF-401-NA, R&D Systems, 1:500), Nlrp3 (AG-20B-0014, Adipogen, 1:500), Caspase-1 (AG-20B-0042, Adipogen, 1:500) and β-Actin (#4967, Cell Signaling,1:2000) overnight at 4 °C.

Techniques: Western Blot, Derivative Assay, Comparison

Journal: Frontiers in Immunology

Article Title: TGFβ1 attenuates microglial IL1β release through inhibition of NLRP3 inflammasome priming

doi: 10.3389/fimmu.2025.1623643

Figure Lengend Snippet: TGFβ1 inhibits LPS-induced expression of Nlrp3 , Casp1 , Il18 , and Il1b in primary microglia. Microglia were treated with TGFβ1 (5 ng/ml), LPS (1 µg/ml) or the combination of both factors for 6 h and RNA and total proteins were isolated. TGFβ1 significantly inhibits LPS-induced upregulation of Nlrp3 (B) , Casp1 (B) , Il18 (C) , and Il1b (D) in primary microglia. TGFβ1 abrogates LPS-induced increases in NLRP3, CASP1, and IL1β protein levels in microglia. Representative western blot images are depicted (E) . Quantifications after normalization using β-Actin are presented for NLRP3 (F) , CASP1 (G) , and IL1β (H) . Data are given as means ± SEM from at least three independent experiments. P-values derived from one-way ANOVA followed by Tukey´s multiple comparison tests are * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: They were later incubated with primary antibodies against Il1β (AF-401-NA, R&D Systems, 1:500), Nlrp3 (AG-20B-0014, Adipogen, 1:500), Caspase-1 (AG-20B-0042, Adipogen, 1:500) and β-Actin (#4967, Cell Signaling,1:2000) overnight at 4 °C.

Techniques: Expressing, Isolation, Western Blot, Derivative Assay, Comparison

Journal: Frontiers in Immunology

Article Title: TGFβ1 attenuates microglial IL1β release through inhibition of NLRP3 inflammasome priming

doi: 10.3389/fimmu.2025.1623643

Figure Lengend Snippet: TGFβ1 reduces release of inflammasome cleaved IL1β from primary microglia. Primary microglia were treated with TGFβ1 (5 ng/ml), LPS (1 µg/ml) or the combination of both factors for 6 h and inflammasome assembly was triggered using Nigericin (1.34mM/ul) treatment for additional 2 h. Proteins were isolated from whole cell lysates (WCL) and supernatants and pro- IL1β and cleaved IL1β was visualized. Representative western blot images are depicted (A) . Quantifications of pro-IL1β after normalization using β-Actin is presented (B) . Total levels of cleaved and bioactive IL1β in supernatants were quantified using ELISA. Quantifications of released IL1β are given (C) . Data are given as means ± SEM from at least three independent experiments. P-values derived from one-way ANOVA followed by Tukey´s multiple comparison tests are ** p < 0.01, and **** p < 0.0001.

Article Snippet: They were later incubated with primary antibodies against Il1β (AF-401-NA, R&D Systems, 1:500), Nlrp3 (AG-20B-0014, Adipogen, 1:500), Caspase-1 (AG-20B-0042, Adipogen, 1:500) and β-Actin (#4967, Cell Signaling,1:2000) overnight at 4 °C.

Techniques: Isolation, Western Blot, Enzyme-linked Immunosorbent Assay, Derivative Assay, Comparison