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tnf α neutralizing antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc tnf α neutralizing antibody
    Immunomodulatory exhaustion-like phenotype of MSCs under <t>prolonged</t> <t>TNF-α</t> stimulation. (A) Schematic representation of indirect co-culture system. MSCs were repeatedly exposed to inflammatory stimulation by transferring them to freshly polarized M1 macrophages every 12 h using Transwell inserts. (B) Time-course analysis of immunoregulatory gene expression ( Tgf-β , Il-10 , and Fasl ) in MSCs during co-culture. Tgf-β expression peaked at 12 h, while Il-10 and Fasl were transiently upregulated and then declined. (C) Progressive increase in the expression of immune checkpoint genes ( Pd-1 and Ctla-4 ) over time. (D) Representative immunofluorescence images showing co-expression of PDGFRα and PD-1 in MSCs after 48 h of co-culture with M1 macrophages (Scale bar = 20 µm). Images are representative of independent experiments and are shown to illustrate the spatial distribution and co-expression of PDGFRα and PD-1, rather than serving as quantitative evidence. (E) Increased Tnf-α and iNos expression in the bone marrow following intraperitoneal injection of LPS-PG (5 mg/kg) for 7 days. (F) Immunohistochemistry of bone marrow showing decreased PDGFRα + TGF-β + cells and increased PDGFRα + PD-1 + cells in LPS-treated mice compared to untreated controls (Scale bar = 100 µm). Quantitative analysis was performed at this magnification, which allowed for the reliable identification of PD-1–positive MSC-like cells. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
    Tnf α Neutralizing Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1786 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/tnf+%CE%B1+neutralizing+antibody/TNF-alpha+Antibody/pmc13002830-35-11-14
    Average 96 stars, based on 1786 article reviews
    tnf α neutralizing antibody - by Bioz Stars, 2026-09
    96/100 stars

    Images

    1) Product Images from "Prolonged TNF-α stimulation induces a PD-1–associated exhaustion-like phenotype in mesenchymal stromal cells"

    Article Title: Prolonged TNF-α stimulation induces a PD-1–associated exhaustion-like phenotype in mesenchymal stromal cells

    Journal: Frontiers in Cell and Developmental Biology

    doi: 10.3389/fcell.2026.1680076

    Immunomodulatory exhaustion-like phenotype of MSCs under prolonged TNF-α stimulation. (A) Schematic representation of indirect co-culture system. MSCs were repeatedly exposed to inflammatory stimulation by transferring them to freshly polarized M1 macrophages every 12 h using Transwell inserts. (B) Time-course analysis of immunoregulatory gene expression ( Tgf-β , Il-10 , and Fasl ) in MSCs during co-culture. Tgf-β expression peaked at 12 h, while Il-10 and Fasl were transiently upregulated and then declined. (C) Progressive increase in the expression of immune checkpoint genes ( Pd-1 and Ctla-4 ) over time. (D) Representative immunofluorescence images showing co-expression of PDGFRα and PD-1 in MSCs after 48 h of co-culture with M1 macrophages (Scale bar = 20 µm). Images are representative of independent experiments and are shown to illustrate the spatial distribution and co-expression of PDGFRα and PD-1, rather than serving as quantitative evidence. (E) Increased Tnf-α and iNos expression in the bone marrow following intraperitoneal injection of LPS-PG (5 mg/kg) for 7 days. (F) Immunohistochemistry of bone marrow showing decreased PDGFRα + TGF-β + cells and increased PDGFRα + PD-1 + cells in LPS-treated mice compared to untreated controls (Scale bar = 100 µm). Quantitative analysis was performed at this magnification, which allowed for the reliable identification of PD-1–positive MSC-like cells. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
    Figure Legend Snippet: Immunomodulatory exhaustion-like phenotype of MSCs under prolonged TNF-α stimulation. (A) Schematic representation of indirect co-culture system. MSCs were repeatedly exposed to inflammatory stimulation by transferring them to freshly polarized M1 macrophages every 12 h using Transwell inserts. (B) Time-course analysis of immunoregulatory gene expression ( Tgf-β , Il-10 , and Fasl ) in MSCs during co-culture. Tgf-β expression peaked at 12 h, while Il-10 and Fasl were transiently upregulated and then declined. (C) Progressive increase in the expression of immune checkpoint genes ( Pd-1 and Ctla-4 ) over time. (D) Representative immunofluorescence images showing co-expression of PDGFRα and PD-1 in MSCs after 48 h of co-culture with M1 macrophages (Scale bar = 20 µm). Images are representative of independent experiments and are shown to illustrate the spatial distribution and co-expression of PDGFRα and PD-1, rather than serving as quantitative evidence. (E) Increased Tnf-α and iNos expression in the bone marrow following intraperitoneal injection of LPS-PG (5 mg/kg) for 7 days. (F) Immunohistochemistry of bone marrow showing decreased PDGFRα + TGF-β + cells and increased PDGFRα + PD-1 + cells in LPS-treated mice compared to untreated controls (Scale bar = 100 µm). Quantitative analysis was performed at this magnification, which allowed for the reliable identification of PD-1–positive MSC-like cells. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Techniques Used: Co-Culture Assay, Transferring, Gene Expression, Expressing, Immunofluorescence, Injection, Immunohistochemistry, Derivative Assay

    MSC dysfunction persists after withdrawal of TNF-α stimulation. (A) Schematic of the withdrawal experiment: MSCs were stimulated with TNF-α for 36 h, followed by culture in TNF-α–free medium. (B) Expression of Tgf-β, Il-10, and Fasl remained suppressed after TNF-α withdrawal. (C) Pd-1 and Ctla-4 expression declined slightly but did not return to baseline. (D) Expression of TNF-α signaling genes (Tradd, Ikkα, and Nf-κb) decreased gradually but remained elevated. (E) Apoptosis-related genes ( Casp3 and Bax ) remained upregulated, indicating sustained cellular stress associated with persistent MSC dysfunction. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4−5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
    Figure Legend Snippet: MSC dysfunction persists after withdrawal of TNF-α stimulation. (A) Schematic of the withdrawal experiment: MSCs were stimulated with TNF-α for 36 h, followed by culture in TNF-α–free medium. (B) Expression of Tgf-β, Il-10, and Fasl remained suppressed after TNF-α withdrawal. (C) Pd-1 and Ctla-4 expression declined slightly but did not return to baseline. (D) Expression of TNF-α signaling genes (Tradd, Ikkα, and Nf-κb) decreased gradually but remained elevated. (E) Apoptosis-related genes ( Casp3 and Bax ) remained upregulated, indicating sustained cellular stress associated with persistent MSC dysfunction. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4−5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Techniques Used: Expressing, Derivative Assay

    TNF-α neutralization partially restores immunoregulatory gene expression in MSCs. (A) Expression of Tgf-β , Il-10 , and Fasl in MSCs co-cultured with M1 macrophages for 12 and 48 h, with or without TNF-α neutralizing antibody. Neutralization inhibited early upregulation (12 h) and prevented the subsequent downregulation observed at 48 h. (B) TNF-α neutralization suppressed the late-phase induction of Pd-1 and Ctla-4 . (C) Expression of TNF-α signaling-related genes ( Tradd , Ikkα , and Nf-κb ) was also reduced after antibody treatment. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
    Figure Legend Snippet: TNF-α neutralization partially restores immunoregulatory gene expression in MSCs. (A) Expression of Tgf-β , Il-10 , and Fasl in MSCs co-cultured with M1 macrophages for 12 and 48 h, with or without TNF-α neutralizing antibody. Neutralization inhibited early upregulation (12 h) and prevented the subsequent downregulation observed at 48 h. (B) TNF-α neutralization suppressed the late-phase induction of Pd-1 and Ctla-4 . (C) Expression of TNF-α signaling-related genes ( Tradd , Ikkα , and Nf-κb ) was also reduced after antibody treatment. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Techniques Used: Neutralization, Gene Expression, Expressing, Cell Culture, Derivative Assay

    Prolonged TNF-α stimulation induces an exhaustion-like phenotype in MSCs. (A) Time-course expression of Tgf-β , Il-10 , and Fasl in MSCs stimulated with TNF-α (10 ng/mL) for up to 48 h. An initial upregulation at the indicated time points was observed, followed by significant downregulation. (B) Immune checkpoint genes ( Pd-1 and Ctla-4 ) were progressively upregulated over time. (C) Immunofluorescence revealed decreased numbers of TGF-β + and IL-10 + MSCs and increased PD-1 + MSCs with prolonged TNF-α stimulation (Scale bar = 20 µm). (D) Expression of Tradd , Ikkα , and Nf-κb remained elevated throughout stimulation. (E) Western blot analysis showed sustained phosphorylation of p65 in MSCs treated with TNF-α. Quantification was performed by densitometric analysis of independent biological replicates (n = 3), each derived from separately prepared MSC cultures. (F) Prolonged TNF-α stimulation led to an increased number of ROS-positive MSCs, indicating elevated intracellular oxidative stress (Scale bar = 100 µm). ROS positivity was defined using a fixed fluorescence intensity threshold, and all samples were processed in parallel, under identical acquisition settings. ROS data are presented as supportive evidence rather than definitive mechanistic proof. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
    Figure Legend Snippet: Prolonged TNF-α stimulation induces an exhaustion-like phenotype in MSCs. (A) Time-course expression of Tgf-β , Il-10 , and Fasl in MSCs stimulated with TNF-α (10 ng/mL) for up to 48 h. An initial upregulation at the indicated time points was observed, followed by significant downregulation. (B) Immune checkpoint genes ( Pd-1 and Ctla-4 ) were progressively upregulated over time. (C) Immunofluorescence revealed decreased numbers of TGF-β + and IL-10 + MSCs and increased PD-1 + MSCs with prolonged TNF-α stimulation (Scale bar = 20 µm). (D) Expression of Tradd , Ikkα , and Nf-κb remained elevated throughout stimulation. (E) Western blot analysis showed sustained phosphorylation of p65 in MSCs treated with TNF-α. Quantification was performed by densitometric analysis of independent biological replicates (n = 3), each derived from separately prepared MSC cultures. (F) Prolonged TNF-α stimulation led to an increased number of ROS-positive MSCs, indicating elevated intracellular oxidative stress (Scale bar = 100 µm). ROS positivity was defined using a fixed fluorescence intensity threshold, and all samples were processed in parallel, under identical acquisition settings. ROS data are presented as supportive evidence rather than definitive mechanistic proof. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Techniques Used: Expressing, Immunofluorescence, Western Blot, Phospho-proteomics, Derivative Assay, Fluorescence

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    Activation Assay:

    Article Title: Microglial reprogramming by Hv1 antagonism protects neurons from inflammatory and glutamate toxicity.
    Article Snippet: .. Additionally, 2,2′- [1,2- Ethanediyl bis(nitrilomethylidyne)] bis[6- methoxy- phenol manganese] complex (EUK134; 100 μM; Millipore, #81065), dizocilpine hydrogen maleate (MK801; 10 μM; Millipore, M107), 2- pyridinylmethylamino ethylamine benzenesulfonic acid hydrate sodium salt (ZX1; 5 μM; Strem Co., #07– 0350), TNF- α neutralizing antibody (nTNF- α; 10 μg/ml; Cell signaling, mAb11969, RRID:AB_2797395), or IL- 6 neutralizing antibody (nIL- 6; 5 μg/ml; InvivoGen #mabg- mil6- 3) were co- administered to characterize the participation of neurotoxic molecules resulting from pro- inflammatory activation. ..

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    Control:

    Article Title: IL-1-induced JAK/STAT signaling is antagonized by TGF-β to shape CAF heterogeneity in pancreatic ductal adenocarcinoma.
    Article Snippet: .. Cells were treated with 0.1 or 1 ng/mL mouse (400-ML-005/CF, R&D Systems) or human (200-LA-002/CF, R&D Systems) IL-1α, 1 ng/mL mouse IL-1β (401-ML-005/CF, R&D Systems), 10 ng/mL mouse TNF-α (410-MT-010/CF, R&D Systems), 20 ng/mL mouse (7666-MB-005/CF, R&D systems) or human TGF-β1 (T7039-2UG, Sigma), 500 nM JAK inhibitor AZD1480 (S2162, Selleck Chem), 30 μM IKK-β inhibitor ML102B, 1 μM A83-01 (2939, Tocris Bioscience), 3 μg/mL IL-1α neutralizing antibody (MAB4001, R&D Systems) or an IgG control (400902, Biolegend), 5 μg/mL TNF-α neutralizing antibody (11969S, Cell signaling) or an IgG control (sc-2027, Santa Cruz), 3.4 μg/mL LIF neutralizing antibody (AF449, R&D Systems) or an IgG control (AB-108-C, R&D), 3.8 μg/mL G-CSF (MAB414, R&D Systems) or IL-6 (MAB406, R&D Systems) neutralizing antibodies or an IgG control (MAB005, R&D). ..

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    Concentration Assay:

    Article Title: Prolonged TNF-α stimulation induces a PD-1–associated exhaustion-like phenotype in mesenchymal stromal cells
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    Incubation:

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    Sino Biological tnfα
    Immunomodulatory exhaustion-like phenotype of MSCs under <t>prolonged</t> <t>TNF-α</t> stimulation. (A) Schematic representation of indirect co-culture system. MSCs were repeatedly exposed to inflammatory stimulation by transferring them to freshly polarized M1 macrophages every 12 h using Transwell inserts. (B) Time-course analysis of immunoregulatory gene expression ( Tgf-β , Il-10 , and Fasl ) in MSCs during co-culture. Tgf-β expression peaked at 12 h, while Il-10 and Fasl were transiently upregulated and then declined. (C) Progressive increase in the expression of immune checkpoint genes ( Pd-1 and Ctla-4 ) over time. (D) Representative immunofluorescence images showing co-expression of PDGFRα and PD-1 in MSCs after 48 h of co-culture with M1 macrophages (Scale bar = 20 µm). Images are representative of independent experiments and are shown to illustrate the spatial distribution and co-expression of PDGFRα and PD-1, rather than serving as quantitative evidence. (E) Increased Tnf-α and iNos expression in the bone marrow following intraperitoneal injection of LPS-PG (5 mg/kg) for 7 days. (F) Immunohistochemistry of bone marrow showing decreased PDGFRα + TGF-β + cells and increased PDGFRα + PD-1 + cells in LPS-treated mice compared to untreated controls (Scale bar = 100 µm). Quantitative analysis was performed at this magnification, which allowed for the reliable identification of PD-1–positive MSC-like cells. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
    Tnfα, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    50349-rn023  (sino biological)
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    sino biological 50349-rn023
    Immunomodulatory exhaustion-like phenotype of MSCs under <t>prolonged</t> <t>TNF-α</t> stimulation. (A) Schematic representation of indirect co-culture system. MSCs were repeatedly exposed to inflammatory stimulation by transferring them to freshly polarized M1 macrophages every 12 h using Transwell inserts. (B) Time-course analysis of immunoregulatory gene expression ( Tgf-β , Il-10 , and Fasl ) in MSCs during co-culture. Tgf-β expression peaked at 12 h, while Il-10 and Fasl were transiently upregulated and then declined. (C) Progressive increase in the expression of immune checkpoint genes ( Pd-1 and Ctla-4 ) over time. (D) Representative immunofluorescence images showing co-expression of PDGFRα and PD-1 in MSCs after 48 h of co-culture with M1 macrophages (Scale bar = 20 µm). Images are representative of independent experiments and are shown to illustrate the spatial distribution and co-expression of PDGFRα and PD-1, rather than serving as quantitative evidence. (E) Increased Tnf-α and iNos expression in the bone marrow following intraperitoneal injection of LPS-PG (5 mg/kg) for 7 days. (F) Immunohistochemistry of bone marrow showing decreased PDGFRα + TGF-β + cells and increased PDGFRα + PD-1 + cells in LPS-treated mice compared to untreated controls (Scale bar = 100 µm). Quantitative analysis was performed at this magnification, which allowed for the reliable identification of PD-1–positive MSC-like cells. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
    50349 Rn023, supplied by sino biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/tnf+%CE%B1+neutralizing+antibody/TNF-alpha%2FTNFA%2FTNFSF2+Neutralizing+Antibody/pmc12866087-77-0-4
    Average 94 stars, based on 1 article reviews
    50349-rn023 - by Bioz Stars, 2026-09
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    tnf α  (Sino Biological)
    93
    Sino Biological tnf α
    Immunomodulatory exhaustion-like phenotype of MSCs under <t>prolonged</t> <t>TNF-α</t> stimulation. (A) Schematic representation of indirect co-culture system. MSCs were repeatedly exposed to inflammatory stimulation by transferring them to freshly polarized M1 macrophages every 12 h using Transwell inserts. (B) Time-course analysis of immunoregulatory gene expression ( Tgf-β , Il-10 , and Fasl ) in MSCs during co-culture. Tgf-β expression peaked at 12 h, while Il-10 and Fasl were transiently upregulated and then declined. (C) Progressive increase in the expression of immune checkpoint genes ( Pd-1 and Ctla-4 ) over time. (D) Representative immunofluorescence images showing co-expression of PDGFRα and PD-1 in MSCs after 48 h of co-culture with M1 macrophages (Scale bar = 20 µm). Images are representative of independent experiments and are shown to illustrate the spatial distribution and co-expression of PDGFRα and PD-1, rather than serving as quantitative evidence. (E) Increased Tnf-α and iNos expression in the bone marrow following intraperitoneal injection of LPS-PG (5 mg/kg) for 7 days. (F) Immunohistochemistry of bone marrow showing decreased PDGFRα + TGF-β + cells and increased PDGFRα + PD-1 + cells in LPS-treated mice compared to untreated controls (Scale bar = 100 µm). Quantitative analysis was performed at this magnification, which allowed for the reliable identification of PD-1–positive MSC-like cells. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
    Tnf α, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/tnf+%CE%B1+neutralizing+antibody/TNF-alpha+%2F+TNFA+%2F+TNFSF2+Neutralizing+Antibody/pm41271562-378-48-49
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    Image Search Results


    The miR‐941/ Keap1 / Nrf2 axis promotes malignant phenotypes of proliferation and migration. (A, B) miR‐941 upregulates MMP9 and MMP27 at the mRNA (A) and protein (B) levels in A549 cells. (C and D) miR‐941 promotes cell migration in an Nrf2‐dependent manner. (C) Representative wound healing images in A549 cells transfected with miR‐941 mimic or NC, with or without ML385 (5 µM). Scale bar, 200 µm. (D) Quantification of wound closure. (E and F) miR‐941 upregulates TNF‐α mRNA (E) and protein (F) levels. (G) Pro‐proliferative effect of miR‐941 is mediated by secreted TNF‐α. Cell viability of A549 cells treated with conditioned medium from miR‐941 ‐overexpressing cells, with or without Infliximab. (H and I) Pro‐migratory effect requires intact Keap1/Nrf2 axis. (H) The wound healing assay of cells treated with miR‐941/WT medium, miR‐941/KEAP1 mut medium and miR‐941/NRF2 KO medium. Scale bar, 200 µm. (I) Quantification of wound closure. (J and K) TNF‐α upregulation depends on Keap1/Nrf2 axis, measured by RT‐qPCR (J) and ELISA (K) in Keap1‐mutant and Nrf2‐KO cells. (L and M) Transwell invasion assays in A549 cells transfected with miR‐941 mimic or NC. (L) Representative images. Scale bar, 100 µm. (M) Quantification of migrated cells per field. Error bars represent standard deviation ( n = 3). p ‐values indicated were calculated by Student's t ‐test (unpaired). ns: p > 0.05, * p < 0.05; ** p < 0.01; *** p < 0.001.

    Journal: Clinical and Translational Medicine

    Article Title: miR‐941 in extracellular vesicles confers anlotinib resistance via Keap1/Nrf2 axis and represents a therapeutic target in non‐small cell lung cancer

    doi: 10.1002/ctm2.70721

    Figure Lengend Snippet: The miR‐941/ Keap1 / Nrf2 axis promotes malignant phenotypes of proliferation and migration. (A, B) miR‐941 upregulates MMP9 and MMP27 at the mRNA (A) and protein (B) levels in A549 cells. (C and D) miR‐941 promotes cell migration in an Nrf2‐dependent manner. (C) Representative wound healing images in A549 cells transfected with miR‐941 mimic or NC, with or without ML385 (5 µM). Scale bar, 200 µm. (D) Quantification of wound closure. (E and F) miR‐941 upregulates TNF‐α mRNA (E) and protein (F) levels. (G) Pro‐proliferative effect of miR‐941 is mediated by secreted TNF‐α. Cell viability of A549 cells treated with conditioned medium from miR‐941 ‐overexpressing cells, with or without Infliximab. (H and I) Pro‐migratory effect requires intact Keap1/Nrf2 axis. (H) The wound healing assay of cells treated with miR‐941/WT medium, miR‐941/KEAP1 mut medium and miR‐941/NRF2 KO medium. Scale bar, 200 µm. (I) Quantification of wound closure. (J and K) TNF‐α upregulation depends on Keap1/Nrf2 axis, measured by RT‐qPCR (J) and ELISA (K) in Keap1‐mutant and Nrf2‐KO cells. (L and M) Transwell invasion assays in A549 cells transfected with miR‐941 mimic or NC. (L) Representative images. Scale bar, 100 µm. (M) Quantification of migrated cells per field. Error bars represent standard deviation ( n = 3). p ‐values indicated were calculated by Student's t ‐test (unpaired). ns: p > 0.05, * p < 0.05; ** p < 0.01; *** p < 0.001.

    Article Snippet: For in vitro treatments, the following reagents were used: anlotinib (MCE, Cat#HY‐101416), ML385 (MCE, Cat#HY‐100523; specific Nrf2 inhibitor), the TNF‐α neutralizing antibody infliximab (MCE, Cat#HY‐P9970), and the pan‐caspase inhibitor Z‐VAD‐FMK (MCE, Cat#HY‐16658B, 20 μM, 24 h).

    Techniques: Migration, Transfection, Wound Healing Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Mutagenesis, Standard Deviation

    The miR‐941/ Keap1 / Nrf2 signalling signature is active in anlotinib‐resistant patient tumours. (A) RT‐qPCR analysis showing elevated miR‐941 levels in tumour tissues from patients resistant to anlotinib compared to sensitive patients. (B) Representative Western blots and quantitative results showing downregulation of Keap1 and upregulation of Nrf2 and its downstream targets ( Bcl‐2 , Mcl‐1 , Bcl‐xL , TNF‐α , MMP9 , MMP27 ) in resistant tumours. (C–E) RT‐qPCR analysis confirming upregulation of the mRNA levels of (C) anti‐apoptotic genes ( Bcl‐2 , Mcl‐1 , Bcl‐xL ), (D) TNF‐α , and (E) MMPs ( MMP9 , MMP27 ) in resistant versus sensitive patient tumours. (F, G) Kaplan–Meier survival curves showing (F) progression‐free survival (PFS) and (G) overall survival (OS) of anlotinib‐treated NSCLC patients stratified by tumour miR‐941 expression (high vs. low, based on median expression value). Data are presented as mean ± SEM. p ‐values indicated were calculated by Mann–Whitney U tests. ns: p > 0.05, * p < 0.05, ** p < 0.01.

    Journal: Clinical and Translational Medicine

    Article Title: miR‐941 in extracellular vesicles confers anlotinib resistance via Keap1/Nrf2 axis and represents a therapeutic target in non‐small cell lung cancer

    doi: 10.1002/ctm2.70721

    Figure Lengend Snippet: The miR‐941/ Keap1 / Nrf2 signalling signature is active in anlotinib‐resistant patient tumours. (A) RT‐qPCR analysis showing elevated miR‐941 levels in tumour tissues from patients resistant to anlotinib compared to sensitive patients. (B) Representative Western blots and quantitative results showing downregulation of Keap1 and upregulation of Nrf2 and its downstream targets ( Bcl‐2 , Mcl‐1 , Bcl‐xL , TNF‐α , MMP9 , MMP27 ) in resistant tumours. (C–E) RT‐qPCR analysis confirming upregulation of the mRNA levels of (C) anti‐apoptotic genes ( Bcl‐2 , Mcl‐1 , Bcl‐xL ), (D) TNF‐α , and (E) MMPs ( MMP9 , MMP27 ) in resistant versus sensitive patient tumours. (F, G) Kaplan–Meier survival curves showing (F) progression‐free survival (PFS) and (G) overall survival (OS) of anlotinib‐treated NSCLC patients stratified by tumour miR‐941 expression (high vs. low, based on median expression value). Data are presented as mean ± SEM. p ‐values indicated were calculated by Mann–Whitney U tests. ns: p > 0.05, * p < 0.05, ** p < 0.01.

    Article Snippet: For in vitro treatments, the following reagents were used: anlotinib (MCE, Cat#HY‐101416), ML385 (MCE, Cat#HY‐100523; specific Nrf2 inhibitor), the TNF‐α neutralizing antibody infliximab (MCE, Cat#HY‐P9970), and the pan‐caspase inhibitor Z‐VAD‐FMK (MCE, Cat#HY‐16658B, 20 μM, 24 h).

    Techniques: Quantitative RT-PCR, Western Blot, Expressing, MANN-WHITNEY

    Immunomodulatory exhaustion-like phenotype of MSCs under prolonged TNF-α stimulation. (A) Schematic representation of indirect co-culture system. MSCs were repeatedly exposed to inflammatory stimulation by transferring them to freshly polarized M1 macrophages every 12 h using Transwell inserts. (B) Time-course analysis of immunoregulatory gene expression ( Tgf-β , Il-10 , and Fasl ) in MSCs during co-culture. Tgf-β expression peaked at 12 h, while Il-10 and Fasl were transiently upregulated and then declined. (C) Progressive increase in the expression of immune checkpoint genes ( Pd-1 and Ctla-4 ) over time. (D) Representative immunofluorescence images showing co-expression of PDGFRα and PD-1 in MSCs after 48 h of co-culture with M1 macrophages (Scale bar = 20 µm). Images are representative of independent experiments and are shown to illustrate the spatial distribution and co-expression of PDGFRα and PD-1, rather than serving as quantitative evidence. (E) Increased Tnf-α and iNos expression in the bone marrow following intraperitoneal injection of LPS-PG (5 mg/kg) for 7 days. (F) Immunohistochemistry of bone marrow showing decreased PDGFRα + TGF-β + cells and increased PDGFRα + PD-1 + cells in LPS-treated mice compared to untreated controls (Scale bar = 100 µm). Quantitative analysis was performed at this magnification, which allowed for the reliable identification of PD-1–positive MSC-like cells. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Prolonged TNF-α stimulation induces a PD-1–associated exhaustion-like phenotype in mesenchymal stromal cells

    doi: 10.3389/fcell.2026.1680076

    Figure Lengend Snippet: Immunomodulatory exhaustion-like phenotype of MSCs under prolonged TNF-α stimulation. (A) Schematic representation of indirect co-culture system. MSCs were repeatedly exposed to inflammatory stimulation by transferring them to freshly polarized M1 macrophages every 12 h using Transwell inserts. (B) Time-course analysis of immunoregulatory gene expression ( Tgf-β , Il-10 , and Fasl ) in MSCs during co-culture. Tgf-β expression peaked at 12 h, while Il-10 and Fasl were transiently upregulated and then declined. (C) Progressive increase in the expression of immune checkpoint genes ( Pd-1 and Ctla-4 ) over time. (D) Representative immunofluorescence images showing co-expression of PDGFRα and PD-1 in MSCs after 48 h of co-culture with M1 macrophages (Scale bar = 20 µm). Images are representative of independent experiments and are shown to illustrate the spatial distribution and co-expression of PDGFRα and PD-1, rather than serving as quantitative evidence. (E) Increased Tnf-α and iNos expression in the bone marrow following intraperitoneal injection of LPS-PG (5 mg/kg) for 7 days. (F) Immunohistochemistry of bone marrow showing decreased PDGFRα + TGF-β + cells and increased PDGFRα + PD-1 + cells in LPS-treated mice compared to untreated controls (Scale bar = 100 µm). Quantitative analysis was performed at this magnification, which allowed for the reliable identification of PD-1–positive MSC-like cells. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Article Snippet: To assess the role of TNF-α secreted by M1 macrophages, a TNF-α neutralizing antibody (Cell Signaling Technology, MA, United States) was added to the MSC culture medium at a final concentration of 1 μg/mL during co-culture experiment.

    Techniques: Co-Culture Assay, Transferring, Gene Expression, Expressing, Immunofluorescence, Injection, Immunohistochemistry, Derivative Assay

    MSC dysfunction persists after withdrawal of TNF-α stimulation. (A) Schematic of the withdrawal experiment: MSCs were stimulated with TNF-α for 36 h, followed by culture in TNF-α–free medium. (B) Expression of Tgf-β, Il-10, and Fasl remained suppressed after TNF-α withdrawal. (C) Pd-1 and Ctla-4 expression declined slightly but did not return to baseline. (D) Expression of TNF-α signaling genes (Tradd, Ikkα, and Nf-κb) decreased gradually but remained elevated. (E) Apoptosis-related genes ( Casp3 and Bax ) remained upregulated, indicating sustained cellular stress associated with persistent MSC dysfunction. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4−5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Prolonged TNF-α stimulation induces a PD-1–associated exhaustion-like phenotype in mesenchymal stromal cells

    doi: 10.3389/fcell.2026.1680076

    Figure Lengend Snippet: MSC dysfunction persists after withdrawal of TNF-α stimulation. (A) Schematic of the withdrawal experiment: MSCs were stimulated with TNF-α for 36 h, followed by culture in TNF-α–free medium. (B) Expression of Tgf-β, Il-10, and Fasl remained suppressed after TNF-α withdrawal. (C) Pd-1 and Ctla-4 expression declined slightly but did not return to baseline. (D) Expression of TNF-α signaling genes (Tradd, Ikkα, and Nf-κb) decreased gradually but remained elevated. (E) Apoptosis-related genes ( Casp3 and Bax ) remained upregulated, indicating sustained cellular stress associated with persistent MSC dysfunction. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4−5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Article Snippet: To assess the role of TNF-α secreted by M1 macrophages, a TNF-α neutralizing antibody (Cell Signaling Technology, MA, United States) was added to the MSC culture medium at a final concentration of 1 μg/mL during co-culture experiment.

    Techniques: Expressing, Derivative Assay

    TNF-α neutralization partially restores immunoregulatory gene expression in MSCs. (A) Expression of Tgf-β , Il-10 , and Fasl in MSCs co-cultured with M1 macrophages for 12 and 48 h, with or without TNF-α neutralizing antibody. Neutralization inhibited early upregulation (12 h) and prevented the subsequent downregulation observed at 48 h. (B) TNF-α neutralization suppressed the late-phase induction of Pd-1 and Ctla-4 . (C) Expression of TNF-α signaling-related genes ( Tradd , Ikkα , and Nf-κb ) was also reduced after antibody treatment. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Prolonged TNF-α stimulation induces a PD-1–associated exhaustion-like phenotype in mesenchymal stromal cells

    doi: 10.3389/fcell.2026.1680076

    Figure Lengend Snippet: TNF-α neutralization partially restores immunoregulatory gene expression in MSCs. (A) Expression of Tgf-β , Il-10 , and Fasl in MSCs co-cultured with M1 macrophages for 12 and 48 h, with or without TNF-α neutralizing antibody. Neutralization inhibited early upregulation (12 h) and prevented the subsequent downregulation observed at 48 h. (B) TNF-α neutralization suppressed the late-phase induction of Pd-1 and Ctla-4 . (C) Expression of TNF-α signaling-related genes ( Tradd , Ikkα , and Nf-κb ) was also reduced after antibody treatment. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Article Snippet: To assess the role of TNF-α secreted by M1 macrophages, a TNF-α neutralizing antibody (Cell Signaling Technology, MA, United States) was added to the MSC culture medium at a final concentration of 1 μg/mL during co-culture experiment.

    Techniques: Neutralization, Gene Expression, Expressing, Cell Culture, Derivative Assay

    Prolonged TNF-α stimulation induces an exhaustion-like phenotype in MSCs. (A) Time-course expression of Tgf-β , Il-10 , and Fasl in MSCs stimulated with TNF-α (10 ng/mL) for up to 48 h. An initial upregulation at the indicated time points was observed, followed by significant downregulation. (B) Immune checkpoint genes ( Pd-1 and Ctla-4 ) were progressively upregulated over time. (C) Immunofluorescence revealed decreased numbers of TGF-β + and IL-10 + MSCs and increased PD-1 + MSCs with prolonged TNF-α stimulation (Scale bar = 20 µm). (D) Expression of Tradd , Ikkα , and Nf-κb remained elevated throughout stimulation. (E) Western blot analysis showed sustained phosphorylation of p65 in MSCs treated with TNF-α. Quantification was performed by densitometric analysis of independent biological replicates (n = 3), each derived from separately prepared MSC cultures. (F) Prolonged TNF-α stimulation led to an increased number of ROS-positive MSCs, indicating elevated intracellular oxidative stress (Scale bar = 100 µm). ROS positivity was defined using a fixed fluorescence intensity threshold, and all samples were processed in parallel, under identical acquisition settings. ROS data are presented as supportive evidence rather than definitive mechanistic proof. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Prolonged TNF-α stimulation induces a PD-1–associated exhaustion-like phenotype in mesenchymal stromal cells

    doi: 10.3389/fcell.2026.1680076

    Figure Lengend Snippet: Prolonged TNF-α stimulation induces an exhaustion-like phenotype in MSCs. (A) Time-course expression of Tgf-β , Il-10 , and Fasl in MSCs stimulated with TNF-α (10 ng/mL) for up to 48 h. An initial upregulation at the indicated time points was observed, followed by significant downregulation. (B) Immune checkpoint genes ( Pd-1 and Ctla-4 ) were progressively upregulated over time. (C) Immunofluorescence revealed decreased numbers of TGF-β + and IL-10 + MSCs and increased PD-1 + MSCs with prolonged TNF-α stimulation (Scale bar = 20 µm). (D) Expression of Tradd , Ikkα , and Nf-κb remained elevated throughout stimulation. (E) Western blot analysis showed sustained phosphorylation of p65 in MSCs treated with TNF-α. Quantification was performed by densitometric analysis of independent biological replicates (n = 3), each derived from separately prepared MSC cultures. (F) Prolonged TNF-α stimulation led to an increased number of ROS-positive MSCs, indicating elevated intracellular oxidative stress (Scale bar = 100 µm). ROS positivity was defined using a fixed fluorescence intensity threshold, and all samples were processed in parallel, under identical acquisition settings. ROS data are presented as supportive evidence rather than definitive mechanistic proof. Data represent mean ± SD of independent biological replicates derived from separately prepared MSC cultures (n = 4–5). One-way ANOVA with Tukey’s post-hoc test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Article Snippet: To assess the role of TNF-α secreted by M1 macrophages, a TNF-α neutralizing antibody (Cell Signaling Technology, MA, United States) was added to the MSC culture medium at a final concentration of 1 μg/mL during co-culture experiment.

    Techniques: Expressing, Immunofluorescence, Western Blot, Phospho-proteomics, Derivative Assay, Fluorescence