Journal: Bioactive Materials

Article Title: Injection site dictates the immune response to a biodegradable polymer and corresponding collagen regeneration

doi: 10.1016/j.bioactmat.2026.04.004

Figure Lengend Snippet: Denser ID tissue architecture leads to greater DAMPs release upon T-gel injection and enhanced macrophage recruitment to the implantation site. (A) Schematic diagram depicting the proposed mechanism: cell damage caused by T-gel injection releases DAMPs (e.g., HMGB1, HSP70, dsDNA and histone DNA), which activate resident macrophages via the DAMPs–TLRs–MyD88–NF-κB signaling axis, promoting secretion of chemokines such as C-C motif chemokine ligands (CCLs) and recruiting additional macrophages/monocytes from local and distal tissues to the injection site. TLRs, Toll-like receptors; MyD88, Myeloid differentiation primary response protein 88; NF-κB, Nuclear factor kappa-light-chain-enhancer of activated B cells. (B) Comparison of maximum pushing force (N) required for ID vs. SC T-gel injection ( n = 3, data represent mean ± s.d.). ∗∗∗∗P < 0.0001. (C) Immunofluorescence analysis of HMGB1 localization and HSP70 expression in tissues one day after ID or SC T-gel injection ( n = 3, data represent mean ± s.d.). ∗∗P < 0.01. (D) Immunofluorescence staining showing nuclear translocation of NF-κB p65 in macrophages (CD68 + cells) one day after ID or SC T-gel injection ( n = 3, data represent mean ± s.d.). ∗∗P < 0.01. (E) RNA-seq analysis of macrophage-related chemokine expression in tissues after ID or SC T-gel injection. “SC ctrl” and “ID ctrl” denote subcutaneous and intradermal injection of normal saline, respectively. Heatmaps show expression levels (FPKM) for each sample. CCL1 and CCL6 did not show significant differences between any groups and are therefore not displayed. (F) Immunofluorescence quantification of CD68 + cells in tissues 7 days after ID or SC T-gel injection ( n = 3, data represent mean ± s.d.). ∗P < 0.05. (G) GO functional analysis of macrophage-associated terms from RNA-seq data after ID or SC T-gel injection.

Article Snippet: After antigen retrieval and blocking, sections were incubated overnight at 4 °C with primary antibodies targeting vimentin, CD68 (ABclonal, A20803), CD31 (R&D SYSTEMS, AF3628), HMGB1 (Cell Signaling Technology, 3935), HSP70 (ABclonal, A23457), NF-κB p65 (ABclonal, A19653), CD206 (Cell Signaling Technology, 24595), and FAPα (ABclonal, A23789 ).

Techniques: Injection, Comparison, Immunofluorescence, Expressing, Staining, Translocation Assay, RNA Sequencing, Saline, Functional Assay

Journal: International Journal of Molecular Sciences

Article Title: Colocasia esculenta Corm Extract Attenuates Ethanol-Induced Gastric Ulcer by Suppressing NF-κB Expression: An Integrated Network Pharmacology Approach and In Vivo Validation

doi: 10.3390/ijms27177693

Figure Lengend Snippet: ( a ) Representative Western blot of NF-κB p65 expression. Representative Western blot bands showing the expression of NF-κB p65 protein in gastric tissues from the different experimental groups following ethanol-induced gastric injury; ( b ) Quantification of NF-κB p65 protein expression. β-Actin was used as the internal loading control. Note: 1, Normal; 2, Negative; 3, Sucralfate; 4, Quercetin; 5, EECE 200 mg/kg; 6, EECE 400 mg/kg; 7, EECE 800 mg/kg.

Article Snippet: Membranes were incubated overnight at 4 °C with primary antibodies against NF-κB p65 (ABclonal Technology, Wuhan, China; Cat. No. A19653, 1:1000) and β-actin (Minneapolis, MN, USA; Cat. No. MAB8529, 1:1000), the membranes were incubated with goat anti-mouse IRDye 680RD and goat anti-rabbit IRDye 800CW secondary antibodies (LI-COR Biosciences, Lincoln, NE, USA; Cat. Nos.

Techniques: Western Blot, Expressing, Control

Journal: Frontiers in Veterinary Science

Article Title: Anti-inflammatory, analgesic, and anti-immune-mediated polyarthritis activities of topically applied polarity fractions of Clematis florida var. Plena and their underlying mechanisms

doi: 10.3389/fvets.2026.1846520

Figure Lengend Snippet: Effect of C. florida fractions on the phosphorylation of ERK1/2 and NF-κB p65 in AA rats ( n = 3 per group). (A) Representative Western blot images showing phospho-ERK1/2, total ERK1/2, phospho-NF-κB p65, and total NF-κB p65 in synovial tissues from different experimental groups. (B) Quantification of the phospho-ERK1/2 to total ERK1/2 ratio. (C) Quantification of the phospho-NF-κB p65 to total NF-κB p65 ratio. Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test. ## p < 0.01, ### p < 0.001 vs. control group; * p < 0.05, ** p < 0.01 vs. model group.

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: p-ERK1/2 (Thr202/Tyr204; Cell Signaling Technology, USA, #3033S, 1:4000), total ERK1/2 (Cell Signaling Technology, USA, #4695S, 1:1000), NF-κB p65 (phospho-Ser536) (BIOSS, China, bsm-33117M, 1:1000), and NF-κB p65 (total) (ABclonal, China, A4782, 1:2000).

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Frontiers in Veterinary Science

Article Title: Anti-inflammatory, analgesic, and anti-immune-mediated polyarthritis activities of topically applied polarity fractions of Clematis florida var. Plena and their underlying mechanisms

doi: 10.3389/fvets.2026.1846520

Figure Lengend Snippet: Immunohistochemical staining of NF-κB p65 proteins in AA rats. (A) Immunohistochemical staining of p-NF-κB p65 expression. (B) Immunohistochemical staining of total NF-κB p65 expression. (C) Area of p-NF-κB p65 expression on osseous tissue. (D) Area of p-NF-κB p65 expression on synovial tissue. (E) Area of total NF-κB p65 expression on osseous tissue. (F) Area of total NF-κB p65 expression on synovial tissue ( n = 5). All figures were magnified by 200×. Scale bar: 50 μm. Statistical comparisons were performed using one-way ANOVA followed by Dunnett’s post hoc test. # p < 0.05, ### p < 0.001 vs. Mcontrol group; * p < 0.05, ** * p < 0.001 vs. model group.

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: p-ERK1/2 (Thr202/Tyr204; Cell Signaling Technology, USA, #3033S, 1:4000), total ERK1/2 (Cell Signaling Technology, USA, #4695S, 1:1000), NF-κB p65 (phospho-Ser536) (BIOSS, China, bsm-33117M, 1:1000), and NF-κB p65 (total) (ABclonal, China, A4782, 1:2000).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Frontiers in Microbiology

Article Title: Spermidine protects against ETEC-induced intestinal injury by reshaping the gut microbiota and activating the AhR/IL-22 axis

doi: 10.3389/fmicb.2026.1907051

Figure Lengend Snippet: Spermidine suppresses activation of the TLR4/MyD88/NF-κB signaling pathway in ETEC-infected mice. (A) Representative Western blot images showing the protein expression of TLR4, MyD88, phosphorylated NF-κB p65 (p-p65), total NF-κB p65 (p65), and GAPDH in jejunal tissues. (B–D) Densitometric analyses of TLR4 (B) , MyD88 (C) , and p-p65 (D) protein expression normalized to the corresponding loading controls. Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. ** p < 0.01, *** p < 0.001.

Article Snippet: The primary antibodies used for Western blot were TLR4 (Proteintech, Cat No. 66350-1-Ig, 1:4,000), MyD88 (ABclonal, Cat No. A0980, 1:2,000), p-p65 (ABclonal, Cat No. AP1294, 1:2,000), p65 (ABclonal, Cat No. A19653, 1:5,000), GAPDH (Proteintech, Cat No. 60004-1-Ig, 1:20,000), ZO-1 (Proteintech, Cat No. 21773-1-AP, 1:5,000), occludin (Proteintech, Cat No. 66378-1-Ig, 1:5,000), AhR (Proteintech, Cat No. 67785-1-Ig, 1:2,000), and CYP1A1 (Proteintech, Cat No. 13241-1-AP, 1:1,000).

Techniques: Activation Assay, Infection, Western Blot, Expressing, Comparison

Journal: Frontiers in Microbiology

Article Title: Spermidine protects against ETEC-induced intestinal injury by reshaping the gut microbiota and activating the AhR/IL-22 axis

doi: 10.3389/fmicb.2026.1907051

Figure Lengend Snippet: Spermidine preserves intestinal barrier integrity in ETEC-infected mice. (A) Representative Western blot images showing the protein expression of ZO-1, occludin, and GAPDH in jejunal tissues. (B–C) Densitometric analyses of ZO-1 (B) and occludin (C) protein expression. (D–F) Serum levels of fluorescein isothiocyanate-dextran 4 kDa (FD4) (D) , diamine oxidase (DAO) activity (E) , and D-lactate (F) as indicators of intestinal permeability and barrier function. Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. ** p < 0.01, *** p < 0.001.

Article Snippet: The primary antibodies used for Western blot were TLR4 (Proteintech, Cat No. 66350-1-Ig, 1:4,000), MyD88 (ABclonal, Cat No. A0980, 1:2,000), p-p65 (ABclonal, Cat No. AP1294, 1:2,000), p65 (ABclonal, Cat No. A19653, 1:5,000), GAPDH (Proteintech, Cat No. 60004-1-Ig, 1:20,000), ZO-1 (Proteintech, Cat No. 21773-1-AP, 1:5,000), occludin (Proteintech, Cat No. 66378-1-Ig, 1:5,000), AhR (Proteintech, Cat No. 67785-1-Ig, 1:2,000), and CYP1A1 (Proteintech, Cat No. 13241-1-AP, 1:1,000).

Techniques: Infection, Western Blot, Expressing, Activity Assay, Permeability, Comparison

Journal: Frontiers in Microbiology

Article Title: Spermidine protects against ETEC-induced intestinal injury by reshaping the gut microbiota and activating the AhR/IL-22 axis

doi: 10.3389/fmicb.2026.1907051

Figure Lengend Snippet: Pharmacological inhibition of AhR abolishes the protective effects of spermidine against ETEC-induced intestinal injury in vivo . (A) Schematic illustration of the experimental design for AhR inhibition using CH-223191 in the ETEC infection model. (B) Survival curves of mice during the experimental period. (C) Changes in body weight following ETEC challenge. (D) Diarrhea scores. (E) Representative hematoxylin and eosin (H&E)-stained sections of the jejunum. Upper panels show low-magnification views (scale bar = 500 μm), and lower panels show corresponding higher-magnification images of the boxed regions (scale bar = 200 μm). (F) Histopathological injury scores. (G) Quantification of the villus-to-crypt (V/C) ratio. (H–K) Serum concentrations of IL-6 (H) , IL-1β (I) , TNF-α (J) , and IL-10 (K) determined by ELISA. (L–N) Assessment of intestinal permeability by serum FITC-dextran 4 kDa (FD4) (L) , diamine oxidase (DAO) activity (M) , and D-lactate (N) . (O) Representative Western blot images showing the expression of AhR, CYP1A1, ZO-1, and occludin in jejunal tissues. (P–S) Densitometric quantification of AhR (P) , CYP1A1 (Q) , ZO-1 (R) , and occludin (S) protein expression normalized to GAPDH. (T) Serum IL-22 levels measured by ELISA. Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001. ns, not significant.

Article Snippet: The primary antibodies used for Western blot were TLR4 (Proteintech, Cat No. 66350-1-Ig, 1:4,000), MyD88 (ABclonal, Cat No. A0980, 1:2,000), p-p65 (ABclonal, Cat No. AP1294, 1:2,000), p65 (ABclonal, Cat No. A19653, 1:5,000), GAPDH (Proteintech, Cat No. 60004-1-Ig, 1:20,000), ZO-1 (Proteintech, Cat No. 21773-1-AP, 1:5,000), occludin (Proteintech, Cat No. 66378-1-Ig, 1:5,000), AhR (Proteintech, Cat No. 67785-1-Ig, 1:2,000), and CYP1A1 (Proteintech, Cat No. 13241-1-AP, 1:1,000).

Techniques: Inhibition, In Vivo, Infection, Staining, Enzyme-linked Immunosorbent Assay, Permeability, Activity Assay, Western Blot, Expressing