foxp3 d2w8e rabbit monoclonal antibody  (Cell Signaling Technology Inc)

Journal: Gut Microbes

Article Title: Targeted intestinal ROS scavenging alleviated rheumatoid arthritis by microbial metabolite kynurenic acid mediated Treg/Th17 balance through AhR pathway activation

doi: 10.1080/19490976.2026.2739669

Figure Lengend Snippet: BPNSs gavage ameliorated arthritis by remodeling Treg/Th17 imbalance. (A) Detailed schematic of the animal experimental procedures of CIA PBS and CIA BPNSs groups. (B) Arthritis scores in CIA mice. (C) Representative photographs and micro-CT images of paws. (D) Quantitative analyses of bone histomorphometry indexes including BMD, BV/TV, Tb.Th, Tb.Sp, and Tb.N in CIA mice. (E) Representative histology images of joints. Scale bar, 100 μm. (F) Histological scores of the joints. (G) Representative TRAP staining images of joints. Scale bar, 100 μm. (H) Quantitative analyses of TRAP staining. (I–N) Flow cytometric analysis and quantification of IL-17A⁺ Th17 cells and FOXP3⁺ Treg cells among CD4⁺ T cells, as well as the Treg/Th17 ratio, in the spleen, MLN and DLN. (O–S) Flow cytometric analysis and quantification of MDSC-like cells, G-MDSC-like cells and M-MDSC-like cells in the spleen. (T) The expression of CD11c, MHCII and CD86 of MDSC-like cells in spleen detected by flow cytometry and statistically analyzed. (U) Immunofluorescence staining of ZO-1 and Occludin in colon tissue. Scale bar, 100 μm. (V) MFI quantification of ZO-1 and Occludin after normalization. (W) Volcano plot of differentially abundant serum inflammatory proteins in CIA BPNSs versus CIA PBS , as determined by a mouse inflammation antibody array. ns, p ≥ 0.05; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: For naïve CD4+ T cells cultured under Treg-polarizing conditions, the following primary antibodies were used: p-mTOR (Ser2448) (Zenbio, Cat. no. 381557, 1:1,000), mTOR (Abmart, Cat. no. T55306F, 1:500), p-p70 S6 kinase (Thr389) (Cell Signaling Technology, Cat. no. 9205, 1:200), p70 S6 kinase (Zenbio, Cat. no. R25650 , 1:1,000), p-AKT (Ser473) (Zenbio, Cat. no. 310021, 1:500), AKT (Zenbio, Cat. no. R23411 , 1:500), p-4E-BP1 (Thr46) (Zenbio, Cat. no. R22929 , 1:1,000), 4E-BP1 (Zenbio, Cat. no. R24197 , 1:500) and FOXP3 (Bioss, Cat. no. bs-10211R, 1:500).

Techniques: Micro-CT, Histology, Staining, Analysis, Expressing, Flow Cytometry, Immunofluorescence, Tissue, Ab Array

Journal: Gut Microbes

Article Title: Targeted intestinal ROS scavenging alleviated rheumatoid arthritis by microbial metabolite kynurenic acid mediated Treg/Th17 balance through AhR pathway activation

doi: 10.1080/19490976.2026.2739669

Figure Lengend Snippet: Gut microbiota depletion led to a reduction in the efficacy of BPNSs. (A) Detailed schematic of the animal experimental procedures of CIA ABX and CIA ABX+BPNSs groups. (B) Arthritis scores in CIA mice. (C) Representative photographs and micro-CT images of paws. (D) Quantitative analyses of bone histomorphometry indexes including BMD, BV/TV, Tb.Th, Tb.Sp, and Tb.N in CIA mice. (E) Representative histology images of joints. Scale bar, 100 μm. (F) Histological scores of joints. (G) Representative TRAP staining images of joints. Scale bar, 100 μm. (H) Quantitative analyses of TRAP staining. (I–L) Flow cytometric analysis and quantification of IL-17A⁺ Th17 cells and FOXP3⁺ Treg cells among CD4⁺ T cells, as well as the Treg/Th17 ratio, in the spleen and MLN. ns, p ≥ 0.05; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: For naïve CD4+ T cells cultured under Treg-polarizing conditions, the following primary antibodies were used: p-mTOR (Ser2448) (Zenbio, Cat. no. 381557, 1:1,000), mTOR (Abmart, Cat. no. T55306F, 1:500), p-p70 S6 kinase (Thr389) (Cell Signaling Technology, Cat. no. 9205, 1:200), p70 S6 kinase (Zenbio, Cat. no. R25650 , 1:1,000), p-AKT (Ser473) (Zenbio, Cat. no. 310021, 1:500), AKT (Zenbio, Cat. no. R23411 , 1:500), p-4E-BP1 (Thr46) (Zenbio, Cat. no. R22929 , 1:1,000), 4E-BP1 (Zenbio, Cat. no. R24197 , 1:500) and FOXP3 (Bioss, Cat. no. bs-10211R, 1:500).

Techniques: Micro-CT, Histology, Staining, Analysis

Journal: Gut Microbes

Article Title: Targeted intestinal ROS scavenging alleviated rheumatoid arthritis by microbial metabolite kynurenic acid mediated Treg/Th17 balance through AhR pathway activation

doi: 10.1080/19490976.2026.2739669

Figure Lengend Snippet: Fecal transplantation from BPNSs-administered CIA mice ameliorated arthritis. (A) Detailed schematic of the animal experimental procedures of CIA FMT-CIA+PBS and CIA FMT-CIA+BPNSs groups. (B) Arthritis scores in CIA mice. (C Representative photographs and micro-CT images of paws. (D) Quantitative analyses of bone histomorphometry indexes including BMD, BV/TV, Tb.Th, Tb.Sp, and Tb.N in CIA mice. (E) Representative histology images of joints. Scale bar, 100 μm. (F) Histological scores of the joints. (G) Representative TRAP staining images of joints. Scale bar, 100 μm. (H) Quantitative analyses of TRAP staining. (I–L) Flow cytometric analysis and quantification of IL-17A⁺ Th17 cells and FOXP3⁺ Treg cells among CD4⁺ T cells, as well as the Treg/Th17 ratio, in the spleen and MLN. ns, p ≥ 0.05; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: For naïve CD4+ T cells cultured under Treg-polarizing conditions, the following primary antibodies were used: p-mTOR (Ser2448) (Zenbio, Cat. no. 381557, 1:1,000), mTOR (Abmart, Cat. no. T55306F, 1:500), p-p70 S6 kinase (Thr389) (Cell Signaling Technology, Cat. no. 9205, 1:200), p70 S6 kinase (Zenbio, Cat. no. R25650 , 1:1,000), p-AKT (Ser473) (Zenbio, Cat. no. 310021, 1:500), AKT (Zenbio, Cat. no. R23411 , 1:500), p-4E-BP1 (Thr46) (Zenbio, Cat. no. R22929 , 1:1,000), 4E-BP1 (Zenbio, Cat. no. R24197 , 1:500) and FOXP3 (Bioss, Cat. no. bs-10211R, 1:500).

Techniques: Transplantation Assay, Micro-CT, Histology, Staining, Analysis

Journal: Gut Microbes

Article Title: Targeted intestinal ROS scavenging alleviated rheumatoid arthritis by microbial metabolite kynurenic acid mediated Treg/Th17 balance through AhR pathway activation

doi: 10.1080/19490976.2026.2739669

Figure Lengend Snippet: KYNA promoted Treg cell differentiation by suppressing mTORC1 activation through the AhR pathway. (A) Predicted binding mode of KYNA within the ligand-binding pocket of AhR. (B) The expression of FOXP3 in naïve CD4⁺ T cells cultured under TGF-β-induced conditions with different concentrations of KYNA, in the presence or absence of CH-223191, was detected by flow cytometry. (C) GSEA of the PI3K/AKT signaling pathway between the control and KYNA-treated groups. (D) Representative Western blot images of AKT, p-AKT, mTOR, p-mTOR, p70 S6K, p-p70 S6K, 4E-BP1, p-4E-BP1, and FOXP3 in the presence or absence of KYNA. (E) Western blot quantification of AKT, p-AKT, mTOR, p-mTOR, p70 S6K, p-p70 S6K, 4E-BP1, p-4E-BP1, and FOXP3 protein levels (normalized to β-actin). ns, p ≥ 0.05; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: For naïve CD4+ T cells cultured under Treg-polarizing conditions, the following primary antibodies were used: p-mTOR (Ser2448) (Zenbio, Cat. no. 381557, 1:1,000), mTOR (Abmart, Cat. no. T55306F, 1:500), p-p70 S6 kinase (Thr389) (Cell Signaling Technology, Cat. no. 9205, 1:200), p70 S6 kinase (Zenbio, Cat. no. R25650 , 1:1,000), p-AKT (Ser473) (Zenbio, Cat. no. 310021, 1:500), AKT (Zenbio, Cat. no. R23411 , 1:500), p-4E-BP1 (Thr46) (Zenbio, Cat. no. R22929 , 1:1,000), 4E-BP1 (Zenbio, Cat. no. R24197 , 1:500) and FOXP3 (Bioss, Cat. no. bs-10211R, 1:500).

Techniques: Cell Differentiation, Activation Assay, Binding Assay, Ligand Binding Assay, Expressing, Cell Culture, Flow Cytometry, Control, Western Blot

Journal: Gut Microbes

Article Title: Targeted intestinal ROS scavenging alleviated rheumatoid arthritis by microbial metabolite kynurenic acid mediated Treg/Th17 balance through AhR pathway activation

doi: 10.1080/19490976.2026.2739669

Figure Lengend Snippet: KYNA exerted anti-rheumatic arthritis effects in vivo through the AhR pathway. (A) Detailed schematic of the animal experimental procedures of CIA PBS , CIA KYNA and CIA KYNA+CH-223191 groups. (B) Arthritis scores in CIA mice. (C) Representative photographs and micro-CT images of paws. (D) Quantitative analyses of bone histomorphometry indexes including BMD, BV/TV, Tb.Th, Tb.Sp, and Tb.N in CIA mice. (E) Representative HE staining images of joints. Scale bar, 100 μm. (F) Histological scores of the joints. (G) Representative TRAP staining images of joints. Scale bar, 100 μm. (H) Quantitative analyses of TRAP staining. (I–L) Flow cytometric analysis and quantification of IL-17A⁺ Th17 cells and FOXP3⁺ Treg cells among CD4⁺ T cells, as well as the Treg/Th17 ratio, in the spleen and MLN. (M–P) (M and O), Representative immunofluorescence staining of ZO-1 and Occludin in colon tissue. Scale bar, 100 μm. (N and P) Quantification of the MFI of ZO-1 and Occludin after normalization. (Q) Representative Western blot images of ZO-1 and Occludin in the presence of KYNA or KYNA + CH-223191 in IEC-6 cells. (R and S) Western blot quantification of ZO-1 and Occludin protein levels (normalized to β-actin). ns, p ≥ 0.05; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: For naïve CD4+ T cells cultured under Treg-polarizing conditions, the following primary antibodies were used: p-mTOR (Ser2448) (Zenbio, Cat. no. 381557, 1:1,000), mTOR (Abmart, Cat. no. T55306F, 1:500), p-p70 S6 kinase (Thr389) (Cell Signaling Technology, Cat. no. 9205, 1:200), p70 S6 kinase (Zenbio, Cat. no. R25650 , 1:1,000), p-AKT (Ser473) (Zenbio, Cat. no. 310021, 1:500), AKT (Zenbio, Cat. no. R23411 , 1:500), p-4E-BP1 (Thr46) (Zenbio, Cat. no. R22929 , 1:1,000), 4E-BP1 (Zenbio, Cat. no. R24197 , 1:500) and FOXP3 (Bioss, Cat. no. bs-10211R, 1:500).

Techniques: In Vivo, Micro-CT, H&E Stain, Staining, Analysis, Immunofluorescence, Tissue, Western Blot

Journal: Gut Microbes

Article Title: Targeted intestinal ROS scavenging alleviated rheumatoid arthritis by microbial metabolite kynurenic acid mediated Treg/Th17 balance through AhR pathway activation

doi: 10.1080/19490976.2026.2739669

Figure Lengend Snippet: Schematic illustration of the protective mechanism of BPNSs against RA progression. The ROS scavenger BPNSs remodeled the gut microbiota, altered KYNA levels, and promoted Treg cell differentiation via the AhR-AKT-mTOR signaling pathway, thereby ameliorating RA progression. Created in https://BioRender.com .

Article Snippet: For naïve CD4+ T cells cultured under Treg-polarizing conditions, the following primary antibodies were used: p-mTOR (Ser2448) (Zenbio, Cat. no. 381557, 1:1,000), mTOR (Abmart, Cat. no. T55306F, 1:500), p-p70 S6 kinase (Thr389) (Cell Signaling Technology, Cat. no. 9205, 1:200), p70 S6 kinase (Zenbio, Cat. no. R25650 , 1:1,000), p-AKT (Ser473) (Zenbio, Cat. no. 310021, 1:500), AKT (Zenbio, Cat. no. R23411 , 1:500), p-4E-BP1 (Thr46) (Zenbio, Cat. no. R22929 , 1:1,000), 4E-BP1 (Zenbio, Cat. no. R24197 , 1:500) and FOXP3 (Bioss, Cat. no. bs-10211R, 1:500).

Techniques: Cell Differentiation

Journal: Research

Article Title: Inverse Vaccine Nanomimics: In situ Antigen-Specific Immune Tolerance Induction for Durable Cardiac Repair Post-Myocardial Infarction

doi: 10.34133/research.1464

Figure Lengend Snippet: Phenotypic changes and inflammatory factor secretion of T cells stimulated by cardiac lysate antigens (CLAs). (A) Coculture protocol for dendritic cells (DCs) and T cells. The tolerogenic DCs (tDCs) present antigens to T cells; meanwhile, programmed death ligand 1 binds to programmed cell death protein 1 on T cells and inhibits T cell activation, promoting the phenotypic conversion of T cells to regulatory T cells (Tregs). Furthermore, due to the lack of activation signals such as CD80 and CD86, some T cells exhibit a weak activation state after receiving antigen presentation, and under the regulation of key factors such as interleukin-10 (IL-10) and transforming growth factor-β (TGF-β), they undergo phenotypic transition to Tregs. Under restimulation, tDCs phagocytose antigens and present them to Tregs. Tregs are rapidly activated, express high levels of Forkhead box P3 (FOXP3), and directly inhibit T cell activation, while secreting large amounts of anti-inflammatory factors. (B) Flow cytometry (FC) results for T cell phenotype. (C) Quantification of T cell phenotypes by FC. (D) T cell phenotype immunofluorescence staining results, where blue represents cell nuclei, red represents CD127, and green represents CD4. (E) Expression of interferon-γ (IFN-γ), IL-2, IL-10, and TGF-β after coculture of DCs and T cells. (F) Western blot analysis of the activation status of the mechanistic target of rapamycin (mTOR)-p70S6K-S6 pathway after treatment with Gel@PPM, Rapa, inverse vaccine nanomimics (IVNMs), and Gel@IVNMs under CLA stimulation. (G) The percentage of phosphorylation of mTOR, p70S6K, and S6 was statistically analyzed based on the gray values of the western blot results (compared to the unphosphorylated proteins). Data are presented as means ± SD, with individual points representing biological replicates. Statistical significance is indicated as ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: After surface staining, cells were fixed and permeabilized with Fixation/Permeabilization Concentrate and Permeabilization Buffer (E-CK-A108, Elabscience, China) according to the manufacturer’s instructions and intracellularly stained with an anti-rat FOXP3 antibody (E-AB-F1351J, Elabscience, China).

Techniques: Activation Assay, Immunopeptidomics, Flow Cytometry, Immunofluorescence, Staining, Expressing, Western Blot, Analysis, Phospho-proteomics

Journal: Research

Article Title: Inverse Vaccine Nanomimics: In situ Antigen-Specific Immune Tolerance Induction for Durable Cardiac Repair Post-Myocardial Infarction

doi: 10.34133/research.1464

Figure Lengend Snippet: Changes in dendritic cell phenotype and regulatory T cell responses in cardiac-draining lymph nodes after myocardial infarction (MI). (A) Representative flow cytometry plots showing CD11c + MHC-II + dendritic cells and programmed death ligand 1 (PD-L1) expression in the Control, Gel@PPM, Gel@PP@Rapa, Gel@OVA-IVNMs, and Gel@IVNMs groups at 7 and 56 d after MI. (B) Quantification of the proportion of PD-L1 + cells within the CD11c + MHC-II + dendritic cell population at 7 and 56 d. (C) Quantification of CD4 + CD25 + FOXP3 + regulatory T cells (Tregs) in lymph node-derived cells from the 5 treatment groups after 48 h of culture under unstimulated Control, OVA, or cardiac lysate antigen (CLA) stimulation at 7 and 56 d. (D) Representative flow cytometry plots of CD25 and Forkhead box P3 (FOXP3) expression within CD4 + T cells in the Gel@OVA-IVNMs and Gel@IVNMs groups following Control, OVA, or CLA stimulation at 7 and 56 d. Data are presented as means ± SD, with individual points representing biological replicates. Statistical significance is indicated as ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: After surface staining, cells were fixed and permeabilized with Fixation/Permeabilization Concentrate and Permeabilization Buffer (E-CK-A108, Elabscience, China) according to the manufacturer’s instructions and intracellularly stained with an anti-rat FOXP3 antibody (E-AB-F1351J, Elabscience, China).

Techniques: Flow Cytometry, Expressing, Control, Derivative Assay