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anti phospho stat4 tyr693  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc anti phospho stat4 tyr693
    Anti Phospho Stat4 Tyr693, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/stat4/Phospho-Stat4+(Tyr693)+Rabbit+mAb/pm41826817-101-69-74
    Average 94 stars, based on 40 article reviews
    anti phospho stat4 tyr693 - by Bioz Stars, 2026-10
    94/100 stars

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    Generated:

    Article Title: Furamidine enhances IL-23-mediated autophagic response through IL-23R-TYK2-STAT3-dependent regulation of intracellular Ca²⁺ level to facilitate mycobacterial clearance in human macrophages.
    Article Snippet: Extended author information available on the last page of the article Abstract Mycobacterium tuberculosis (M. tb), the etiological agent of tuberculosis (TB), subverts host immune responses to establish persistent infection by influencing cytokine production.. An effective and balanced cytokine response is crucial for eliminating the pathogen, whereas disruptions in this balance can lead to increased vulnerability to infection.. Therefore, developing immunomodulatory agents to steer host immunity toward the adequate clearance of M. tb offers a promising strategy for limiting TB.

    Western Blot:

    Article Title: Identification of a JAK–STAT–miR155HG positive feedback loop in regulating natural killer (NK) cells proliferation and effector functions
    Article Snippet: .. The following antibodies were used in Western blot: rabbit monoclonal antibodies (mAbs) against human phospho-Tyr980/981 of JAK3 (5031t), human phospho-Y705 of STAT3 (9145T), STAT3 (4904S), phospho-Y693 of STAT4 (4134S), STAT4 (2653S), phospho-Y694 of STAT5 (9359S), STAT5 (94205S), GAPDH (8884S) from Cell Signalling Technology (CST, Beverly, MA, USA); rabbit monoclonal antibodies (mAbs) against JAK3 (ab45141), phospho-Tyr1022/1023 of JAK1 (ab138005), JAK1 (ab133666) from Abcam Jo urn al Pr -pr oo f 5 (Cambridge, UK). .. The following antibodies were used in flow cytometry analysis: CD45 (#560178, BD Biosciences; #304037, BioLegend), CD56 (#562780, BD Biosciences; #392406, BioLegend; #IM2474, Beckman Coulter, Miami, FL, USA), CD3 (#300316, BioLegend), NKp46 (#331914, BioLegend), CD16 (#302012, BioLegend), CD34 (#343516, BioLegend), CD43 (#343206, BioLegend), IFN-γ (#502530, BioLegend), Ki67 (#350530, BioLegend), granzyme B (#561142, BD Biosciences), perforin (#353314, BioLegend), CD107a (#555801, BD Biosciences), 7-AAD antibodies (#559763, BD Bioscience), NKP30 (#130-112-430, Miltenyi, Bergisch Gladbach Germany), KIR2DL5A (#566330, BD Biosciences).

    Article Title: Identification of a JAK–STAT–miR155HG positive feedback loop in regulating natural killer (NK) cells proliferation and effector functions
    Article Snippet: .. The following antibodies were used in Western blot: rabbit monoclonal antibodies (mAbs) against human phospho-Tyr980/981 of JAK3 (5031t), human phospho-Y705 of STAT3 (9145T), STAT3 (4904S), phospho-Y693 of STAT4 (4134S), STAT4 (2653S), phospho-Y694 of STAT5 (9359S), STAT5 (94205S), GAPDH (8884S) from Cell Signalling Technology (CST, Beverly, MA, USA); rabbit monoclonal antibodies (mAbs) against JAK3 (ab45141), phospho-Tyr1022/1023 of JAK1 (ab138005), JAK1 (ab133666) from Abcam (Cambridge, UK). .. The following antibodies were used in flow cytometry analysis: CD45 (#560178; BD Biosciences; #304037; BioLegend), CD56 (#562780, BD Biosciences; #392406, BioLegend; #IM2474, Beckman Coulter, Miami, FL, USA), CD3 (#300316; BioLegend), NKp46 (#331914; BioLegend), CD16 (#302012; BioLegend), CD34 (#343516; BioLegend), CD43 (#343206; BioLegend), IFN- γ (#502530; BioLegend), Ki67 (#350530; BioLegend), granzyme B (#561142; BD Biosciences), perforin (#353314; BioLegend), CD107a (#555801; BD Biosciences), 7-AAD antibodies (#559763; BD Bioscience), NKP30 (#130-112-430; Miltenyi, Bergisch Gladbach Germany), KIR2DL5A (#566330; BD Biosciences).

    Bioprocessing:

    Article Title: Identification of a JAK–STAT–miR155HG positive feedback loop in regulating natural killer (NK) cells proliferation and effector functions
    Article Snippet: .. The following antibodies were used in Western blot: rabbit monoclonal antibodies (mAbs) against human phospho-Tyr980/981 of JAK3 (5031t), human phospho-Y705 of STAT3 (9145T), STAT3 (4904S), phospho-Y693 of STAT4 (4134S), STAT4 (2653S), phospho-Y694 of STAT5 (9359S), STAT5 (94205S), GAPDH (8884S) from Cell Signalling Technology (CST, Beverly, MA, USA); rabbit monoclonal antibodies (mAbs) against JAK3 (ab45141), phospho-Tyr1022/1023 of JAK1 (ab138005), JAK1 (ab133666) from Abcam Jo urn al Pr -pr oo f 5 (Cambridge, UK). .. The following antibodies were used in flow cytometry analysis: CD45 (#560178, BD Biosciences; #304037, BioLegend), CD56 (#562780, BD Biosciences; #392406, BioLegend; #IM2474, Beckman Coulter, Miami, FL, USA), CD3 (#300316, BioLegend), NKp46 (#331914, BioLegend), CD16 (#302012, BioLegend), CD34 (#343516, BioLegend), CD43 (#343206, BioLegend), IFN-γ (#502530, BioLegend), Ki67 (#350530, BioLegend), granzyme B (#561142, BD Biosciences), perforin (#353314, BioLegend), CD107a (#555801, BD Biosciences), 7-AAD antibodies (#559763, BD Bioscience), NKP30 (#130-112-430, Miltenyi, Bergisch Gladbach Germany), KIR2DL5A (#566330, BD Biosciences).

    Article Title: Identification of a JAK–STAT–miR155HG positive feedback loop in regulating natural killer (NK) cells proliferation and effector functions
    Article Snippet: .. The following antibodies were used in Western blot: rabbit monoclonal antibodies (mAbs) against human phospho-Tyr980/981 of JAK3 (5031t), human phospho-Y705 of STAT3 (9145T), STAT3 (4904S), phospho-Y693 of STAT4 (4134S), STAT4 (2653S), phospho-Y694 of STAT5 (9359S), STAT5 (94205S), GAPDH (8884S) from Cell Signalling Technology (CST, Beverly, MA, USA); rabbit monoclonal antibodies (mAbs) against JAK3 (ab45141), phospho-Tyr1022/1023 of JAK1 (ab138005), JAK1 (ab133666) from Abcam (Cambridge, UK). .. The following antibodies were used in flow cytometry analysis: CD45 (#560178; BD Biosciences; #304037; BioLegend), CD56 (#562780, BD Biosciences; #392406, BioLegend; #IM2474, Beckman Coulter, Miami, FL, USA), CD3 (#300316; BioLegend), NKp46 (#331914; BioLegend), CD16 (#302012; BioLegend), CD34 (#343516; BioLegend), CD43 (#343206; BioLegend), IFN- γ (#502530; BioLegend), Ki67 (#350530; BioLegend), granzyme B (#561142; BD Biosciences), perforin (#353314; BioLegend), CD107a (#555801; BD Biosciences), 7-AAD antibodies (#559763; BD Bioscience), NKP30 (#130-112-430; Miltenyi, Bergisch Gladbach Germany), KIR2DL5A (#566330; BD Biosciences).

    Lysis:

    Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis
    Article Snippet: .. SDS loading buffer (Deeyeebio, 50053) was added to the cell lysis solution, and the proteins (30 μg/lane) were separated by SDS‐PAGE and transferred onto the PVDF membrane, and probed with primary antibodies against phospho‐JAK2 (Tyr1007/1008) (CST, 3771), JAK2 (CST, 3230), phospho‐STAT3 (Ser705) (CST, 9145), phospho‐STAT4 (Tyr693) (CST, 4134), STAT3 (Proteintech, 10253‐2‐ap), STAT4 (CST, 2653), SOCS1 (CST, 55313), SOCS3 (Proteintech, 14025‐1‐ap), and AhR (Proteintech, 67785‐1‐lg), followed by HRP‐conjugated secondary antibodies (Proteintech, SA00001‐1, SA00001‐2). .. Signals were detected via ECL (Deeyeebio, 30208), with GAPDH (Proteintech, 60004‐1‐Ig) as the loading control.

    Membrane:

    Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis
    Article Snippet: .. SDS loading buffer (Deeyeebio, 50053) was added to the cell lysis solution, and the proteins (30 μg/lane) were separated by SDS‐PAGE and transferred onto the PVDF membrane, and probed with primary antibodies against phospho‐JAK2 (Tyr1007/1008) (CST, 3771), JAK2 (CST, 3230), phospho‐STAT3 (Ser705) (CST, 9145), phospho‐STAT4 (Tyr693) (CST, 4134), STAT3 (Proteintech, 10253‐2‐ap), STAT4 (CST, 2653), SOCS1 (CST, 55313), SOCS3 (Proteintech, 14025‐1‐ap), and AhR (Proteintech, 67785‐1‐lg), followed by HRP‐conjugated secondary antibodies (Proteintech, SA00001‐1, SA00001‐2). .. Signals were detected via ECL (Deeyeebio, 30208), with GAPDH (Proteintech, 60004‐1‐Ig) as the loading control.

    Saline:

    Article Title: CD30 knockout attenuates experimental colitis by reducing inflammatory cytokine production.
    Article Snippet: Equal amounts of protein (30 μg) were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Millipore, Burlington, MA, USA) using a Trans-Blot system (Bio-Rad, Hercules, CA, USA). .. Membranes were blocked with 5% non-fat milk (BD Biosciences) in Tris-buffered saline with 0.1% Tween-20 (TBST) for 1 hour at room temperature and incubated AR TIC LE IN PR ES S overnight at 4°C with following primary antibodies (1:1000 dilution): anti-CD30 (Cat. No. AF852; R&D Systems), anti-T-bet (Cat. No. ab91109; Abcam, Cambridge, MA, USA), anti-IFN-γ (Cat. No. ab9657; Abcam), anti-STAT4 (Cat. No. 2653; Cell Signaling Technology, Danvers, MA, USA), anti-phospho-STAT4 (Tyr693) (Cat. No. 4134; Cell Signaling Technology), anti-NF-κB p65 (Cat. No. 8242; Cell Signaling Technology), anti-phospho-NF-κB p65 (Ser536) (Cat. No. 3033; Cell Signaling Technology), anti-IκBα (Cat. No. 4814; Cell Signaling Technology), anti-phospho-IκBα (Ser32) (Cat. No. 2859; Cell Signaling Technology), and anti-β-actin (Cat. No. 4970; Cell Signaling Technology). .. After washing with TBST, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Cat. No. 7074 for anti-rabbit IgG, Cat. No. 7076 for anti-mouse IgG; Cell Signaling Technology, 1:5000 dilution) for 1 hour at room temperature.

    Incubation:

    Article Title: CD30 knockout attenuates experimental colitis by reducing inflammatory cytokine production.
    Article Snippet: Equal amounts of protein (30 μg) were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Millipore, Burlington, MA, USA) using a Trans-Blot system (Bio-Rad, Hercules, CA, USA). .. Membranes were blocked with 5% non-fat milk (BD Biosciences) in Tris-buffered saline with 0.1% Tween-20 (TBST) for 1 hour at room temperature and incubated AR TIC LE IN PR ES S overnight at 4°C with following primary antibodies (1:1000 dilution): anti-CD30 (Cat. No. AF852; R&D Systems), anti-T-bet (Cat. No. ab91109; Abcam, Cambridge, MA, USA), anti-IFN-γ (Cat. No. ab9657; Abcam), anti-STAT4 (Cat. No. 2653; Cell Signaling Technology, Danvers, MA, USA), anti-phospho-STAT4 (Tyr693) (Cat. No. 4134; Cell Signaling Technology), anti-NF-κB p65 (Cat. No. 8242; Cell Signaling Technology), anti-phospho-NF-κB p65 (Ser536) (Cat. No. 3033; Cell Signaling Technology), anti-IκBα (Cat. No. 4814; Cell Signaling Technology), anti-phospho-IκBα (Ser32) (Cat. No. 2859; Cell Signaling Technology), and anti-β-actin (Cat. No. 4970; Cell Signaling Technology). .. After washing with TBST, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Cat. No. 7074 for anti-rabbit IgG, Cat. No. 7076 for anti-mouse IgG; Cell Signaling Technology, 1:5000 dilution) for 1 hour at room temperature.



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    Chemotherapy-induced senescence of nasopharyngeal carcinoma cells upregulates EBI3 to promote IL35 secretion, thereby activating the STAT1-NKG2A signaling axis and affecting NK cell-mediated anti-tumor immunity. A The supernatants of NPC cells without cisplatin treatment, cisplatin-treated WT, and cisplatin-treated sgEBI3 were co-cultured with NK-92MI cells. RT-qPCR was used to analyze the mRNA levels of activating and inhibitory receptors in NK-92MI cells ( n = 3 experiments). B Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. RT-qPCR was used to analyze the mRNA levels of activating and inhibitory receptors in NK-92MI cells ( n = 3 experiments). C Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells. The upper figure is a representative image of flow cytometry, and the lower figure is a statistical chart ( n = 3 experiments). D Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells. The upper figure is a representative image of flow cytometry, and the lower figure is a statistical chart ( n = 3 experiments). E Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells with or without IL-35. RT-qPCR was used to analyze the mRNA levels of IL35R in NK-92MI cells ( n = 3 experiments). F Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells treated with control, r-IL-35, r-IL-35 plus STAT1 inhibitors, and r-IL-35 plus <t>STAT4</t> inhibitors. The left image is a representative flow cytometry image, and the right image is a statistical graph ( n = 3 experiments). G RT-qPCR was used to detect the mRNA levels of NKG2A in NK-92MI cells treated with control, r-IL-35, r-IL-35 + STAT1 inhibitors, and rIL35 + STAT4 inhibitors ( n = 3 experiments). H Schematic diagram of STAT1 binding in the NKG2A promoter region. I ChIP was used to detect the binding of STAT1 to the NKG2A promoter region ( n = 3 experiments). J Schematic diagram of the molecular mechanism by which IL-35 upregulates NKG2A expression in NK cells via activating the STAT1 signaling pathway through its receptor ( A - D , F - H , J ) data are expressed as mean ± SD; B , F , J A two-tailed unpaired t-test was used; A , C , D , G , H A two-tailed unpaired t-test or one-way analysis of variance (ANOVA) and Tukey’s post hoc test was used
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    95
    Cell Signaling Technology Inc antibodies against stat4
    Chemotherapy-induced senescence of nasopharyngeal carcinoma cells upregulates EBI3 to promote IL35 secretion, thereby activating the STAT1-NKG2A signaling axis and affecting NK cell-mediated anti-tumor immunity. A The supernatants of NPC cells without cisplatin treatment, cisplatin-treated WT, and cisplatin-treated sgEBI3 were co-cultured with NK-92MI cells. RT-qPCR was used to analyze the mRNA levels of activating and inhibitory receptors in NK-92MI cells ( n = 3 experiments). B Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. RT-qPCR was used to analyze the mRNA levels of activating and inhibitory receptors in NK-92MI cells ( n = 3 experiments). C Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells. The upper figure is a representative image of flow cytometry, and the lower figure is a statistical chart ( n = 3 experiments). D Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells. The upper figure is a representative image of flow cytometry, and the lower figure is a statistical chart ( n = 3 experiments). E Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells with or without IL-35. RT-qPCR was used to analyze the mRNA levels of IL35R in NK-92MI cells ( n = 3 experiments). F Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells treated with control, r-IL-35, r-IL-35 plus STAT1 inhibitors, and r-IL-35 plus <t>STAT4</t> inhibitors. The left image is a representative flow cytometry image, and the right image is a statistical graph ( n = 3 experiments). G RT-qPCR was used to detect the mRNA levels of NKG2A in NK-92MI cells treated with control, r-IL-35, r-IL-35 + STAT1 inhibitors, and rIL35 + STAT4 inhibitors ( n = 3 experiments). H Schematic diagram of STAT1 binding in the NKG2A promoter region. I ChIP was used to detect the binding of STAT1 to the NKG2A promoter region ( n = 3 experiments). J Schematic diagram of the molecular mechanism by which IL-35 upregulates NKG2A expression in NK cells via activating the STAT1 signaling pathway through its receptor ( A - D , F - H , J ) data are expressed as mean ± SD; B , F , J A two-tailed unpaired t-test was used; A , C , D , G , H A two-tailed unpaired t-test or one-way analysis of variance (ANOVA) and Tukey’s post hoc test was used
    Antibodies Against Stat4, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech 13028-1-ap
    Chemotherapy-induced senescence of nasopharyngeal carcinoma cells upregulates EBI3 to promote IL35 secretion, thereby activating the STAT1-NKG2A signaling axis and affecting NK cell-mediated anti-tumor immunity. A The supernatants of NPC cells without cisplatin treatment, cisplatin-treated WT, and cisplatin-treated sgEBI3 were co-cultured with NK-92MI cells. RT-qPCR was used to analyze the mRNA levels of activating and inhibitory receptors in NK-92MI cells ( n = 3 experiments). B Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. RT-qPCR was used to analyze the mRNA levels of activating and inhibitory receptors in NK-92MI cells ( n = 3 experiments). C Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells. The upper figure is a representative image of flow cytometry, and the lower figure is a statistical chart ( n = 3 experiments). D Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells. The upper figure is a representative image of flow cytometry, and the lower figure is a statistical chart ( n = 3 experiments). E Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells with or without IL-35. RT-qPCR was used to analyze the mRNA levels of IL35R in NK-92MI cells ( n = 3 experiments). F Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells treated with control, r-IL-35, r-IL-35 plus STAT1 inhibitors, and r-IL-35 plus <t>STAT4</t> inhibitors. The left image is a representative flow cytometry image, and the right image is a statistical graph ( n = 3 experiments). G RT-qPCR was used to detect the mRNA levels of NKG2A in NK-92MI cells treated with control, r-IL-35, r-IL-35 + STAT1 inhibitors, and rIL35 + STAT4 inhibitors ( n = 3 experiments). H Schematic diagram of STAT1 binding in the NKG2A promoter region. I ChIP was used to detect the binding of STAT1 to the NKG2A promoter region ( n = 3 experiments). J Schematic diagram of the molecular mechanism by which IL-35 upregulates NKG2A expression in NK cells via activating the STAT1 signaling pathway through its receptor ( A - D , F - H , J ) data are expressed as mean ± SD; B , F , J A two-tailed unpaired t-test was used; A , C , D , G , H A two-tailed unpaired t-test or one-way analysis of variance (ANOVA) and Tukey’s post hoc test was used
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    Image Search Results


    IAAld suppresses Th1 and Th17 cell differentiation in an AhR‐dependent manner by repressing JAK/STAT signaling. (A) KEGG pathway enrichment analysis of the intersectional targets of IAAld and MS. (B) In silico prediction of the binding interaction between IAAld and AhR by molecular docking. The calculated binding energy (BE) is −6.6 kcal/mol (inset: chemical structure of IAAld). (C,D) Flow cytometric quantification and representative plots of CD4 + IFNγ + (Th1) and CD4 + IL‑17A + (Th17) cells under in vitro differentiation conditions across PBS, IAAld (10 µM), and IAAld+siAhR (AhR knockdown by siRNA) groups ( n = 3 per group). (E) Immunofluorescence staining of AhR (green) in Th1 and Th17 cells of each group, and cell nuclei were stained with DAPI (blue). (F) Western blot analysis showing protein levels of AhR, p‑JAK2, JAK2, p‑STAT4, STAT4, p‑STAT3, STAT3, SOCS1, SOCS3, and GAPDH in Th1 (left) and Th17 (right) cells. (G) Quantitative analysis of AhR/GAPDH ratio, p‐JAK2/JAK2 ratio, p‐STAT4/STAT4 ratio, p‐STAT3/STAT3 ratio, SOCS1/GAPDH ratio and SOCS3/GAPDH ratio in each group ( n = 4 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: Advanced Science

    Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

    doi: 10.1002/advs.78358

    Figure Lengend Snippet: IAAld suppresses Th1 and Th17 cell differentiation in an AhR‐dependent manner by repressing JAK/STAT signaling. (A) KEGG pathway enrichment analysis of the intersectional targets of IAAld and MS. (B) In silico prediction of the binding interaction between IAAld and AhR by molecular docking. The calculated binding energy (BE) is −6.6 kcal/mol (inset: chemical structure of IAAld). (C,D) Flow cytometric quantification and representative plots of CD4 + IFNγ + (Th1) and CD4 + IL‑17A + (Th17) cells under in vitro differentiation conditions across PBS, IAAld (10 µM), and IAAld+siAhR (AhR knockdown by siRNA) groups ( n = 3 per group). (E) Immunofluorescence staining of AhR (green) in Th1 and Th17 cells of each group, and cell nuclei were stained with DAPI (blue). (F) Western blot analysis showing protein levels of AhR, p‑JAK2, JAK2, p‑STAT4, STAT4, p‑STAT3, STAT3, SOCS1, SOCS3, and GAPDH in Th1 (left) and Th17 (right) cells. (G) Quantitative analysis of AhR/GAPDH ratio, p‐JAK2/JAK2 ratio, p‐STAT4/STAT4 ratio, p‐STAT3/STAT3 ratio, SOCS1/GAPDH ratio and SOCS3/GAPDH ratio in each group ( n = 4 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: SDS loading buffer (Deeyeebio, 50053) was added to the cell lysis solution, and the proteins (30 μg/lane) were separated by SDS‐PAGE and transferred onto the PVDF membrane, and probed with primary antibodies against phospho‐JAK2 (Tyr1007/1008) (CST, 3771), JAK2 (CST, 3230), phospho‐STAT3 (Ser705) (CST, 9145), phospho‐STAT4 (Tyr693) (CST, 4134), STAT3 (Proteintech, 10253‐2‐ap), STAT4 (CST, 2653), SOCS1 (CST, 55313), SOCS3 (Proteintech, 14025‐1‐ap), and AhR (Proteintech, 67785‐1‐lg), followed by HRP‐conjugated secondary antibodies (Proteintech, SA00001‐1, SA00001‐2).

    Techniques: Cell Differentiation, Analysis, In Silico, Binding Assay, Docking Assay, In Vitro, Knockdown, Immunofluorescence, Staining, Western Blot, Significance Assay

    Graphical abstract. Schematic diagram illustrating the molecular mechanism by which KU ameliorates EAE through the gut microbiota‐IAAld‐AhR axis. KU reshapes gut microbiota composition in EAE mice, specifically increasing the abundance of Akkermansia muciniphila . A. muciniphila facilitates intestinal production of indole‐3‐acetaldehyde (IAAld) and the latter crosses the intestinal barrier to enter peripheral circulation and CNS, where IAAld activates AhR and promotes its nuclear translocation in CD4 + T cells. Once in the nucleus, activated AhR upregulates SOCS1 and SOCS3 expression and suppresses phosphorylation of JAK2 and its downstream targets, STAT4 and STAT3. This cascade restrains CD4 + T cell differentiation into Th1 and Th17 subsets, ultimately mitigating CNS inflammation and demyelination in EAE mice.

    Journal: Advanced Science

    Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

    doi: 10.1002/advs.78358

    Figure Lengend Snippet: Graphical abstract. Schematic diagram illustrating the molecular mechanism by which KU ameliorates EAE through the gut microbiota‐IAAld‐AhR axis. KU reshapes gut microbiota composition in EAE mice, specifically increasing the abundance of Akkermansia muciniphila . A. muciniphila facilitates intestinal production of indole‐3‐acetaldehyde (IAAld) and the latter crosses the intestinal barrier to enter peripheral circulation and CNS, where IAAld activates AhR and promotes its nuclear translocation in CD4 + T cells. Once in the nucleus, activated AhR upregulates SOCS1 and SOCS3 expression and suppresses phosphorylation of JAK2 and its downstream targets, STAT4 and STAT3. This cascade restrains CD4 + T cell differentiation into Th1 and Th17 subsets, ultimately mitigating CNS inflammation and demyelination in EAE mice.

    Article Snippet: SDS loading buffer (Deeyeebio, 50053) was added to the cell lysis solution, and the proteins (30 μg/lane) were separated by SDS‐PAGE and transferred onto the PVDF membrane, and probed with primary antibodies against phospho‐JAK2 (Tyr1007/1008) (CST, 3771), JAK2 (CST, 3230), phospho‐STAT3 (Ser705) (CST, 9145), phospho‐STAT4 (Tyr693) (CST, 4134), STAT3 (Proteintech, 10253‐2‐ap), STAT4 (CST, 2653), SOCS1 (CST, 55313), SOCS3 (Proteintech, 14025‐1‐ap), and AhR (Proteintech, 67785‐1‐lg), followed by HRP‐conjugated secondary antibodies (Proteintech, SA00001‐1, SA00001‐2).

    Techniques: Translocation Assay, Expressing, Phospho-proteomics, Cell Differentiation

    Chemotherapy-induced senescence of nasopharyngeal carcinoma cells upregulates EBI3 to promote IL35 secretion, thereby activating the STAT1-NKG2A signaling axis and affecting NK cell-mediated anti-tumor immunity. A The supernatants of NPC cells without cisplatin treatment, cisplatin-treated WT, and cisplatin-treated sgEBI3 were co-cultured with NK-92MI cells. RT-qPCR was used to analyze the mRNA levels of activating and inhibitory receptors in NK-92MI cells ( n = 3 experiments). B Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. RT-qPCR was used to analyze the mRNA levels of activating and inhibitory receptors in NK-92MI cells ( n = 3 experiments). C Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells. The upper figure is a representative image of flow cytometry, and the lower figure is a statistical chart ( n = 3 experiments). D Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells. The upper figure is a representative image of flow cytometry, and the lower figure is a statistical chart ( n = 3 experiments). E Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells with or without IL-35. RT-qPCR was used to analyze the mRNA levels of IL35R in NK-92MI cells ( n = 3 experiments). F Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells treated with control, r-IL-35, r-IL-35 plus STAT1 inhibitors, and r-IL-35 plus STAT4 inhibitors. The left image is a representative flow cytometry image, and the right image is a statistical graph ( n = 3 experiments). G RT-qPCR was used to detect the mRNA levels of NKG2A in NK-92MI cells treated with control, r-IL-35, r-IL-35 + STAT1 inhibitors, and rIL35 + STAT4 inhibitors ( n = 3 experiments). H Schematic diagram of STAT1 binding in the NKG2A promoter region. I ChIP was used to detect the binding of STAT1 to the NKG2A promoter region ( n = 3 experiments). J Schematic diagram of the molecular mechanism by which IL-35 upregulates NKG2A expression in NK cells via activating the STAT1 signaling pathway through its receptor ( A - D , F - H , J ) data are expressed as mean ± SD; B , F , J A two-tailed unpaired t-test was used; A , C , D , G , H A two-tailed unpaired t-test or one-way analysis of variance (ANOVA) and Tukey’s post hoc test was used

    Journal: Cell Communication and Signaling : CCS

    Article Title: Chemotherapy-induced senescent nasopharyngeal carcinoma cells suppress NK cell-mediated antitumor immunity by upregulating EBI3 via mtDNA-cGAS-STING pathway

    doi: 10.1186/s12964-026-02848-6

    Figure Lengend Snippet: Chemotherapy-induced senescence of nasopharyngeal carcinoma cells upregulates EBI3 to promote IL35 secretion, thereby activating the STAT1-NKG2A signaling axis and affecting NK cell-mediated anti-tumor immunity. A The supernatants of NPC cells without cisplatin treatment, cisplatin-treated WT, and cisplatin-treated sgEBI3 were co-cultured with NK-92MI cells. RT-qPCR was used to analyze the mRNA levels of activating and inhibitory receptors in NK-92MI cells ( n = 3 experiments). B Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. RT-qPCR was used to analyze the mRNA levels of activating and inhibitory receptors in NK-92MI cells ( n = 3 experiments). C Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells. The upper figure is a representative image of flow cytometry, and the lower figure is a statistical chart ( n = 3 experiments). D Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells in the presence or absence of r-IL-35. Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells. The upper figure is a representative image of flow cytometry, and the lower figure is a statistical chart ( n = 3 experiments). E Cisplatin-induced sgEBI3 NPC cells were co-cultured with NK-92MI cells with or without IL-35. RT-qPCR was used to analyze the mRNA levels of IL35R in NK-92MI cells ( n = 3 experiments). F Flow cytometry was used to detect the expression of NKG2A in NK-92MI cells treated with control, r-IL-35, r-IL-35 plus STAT1 inhibitors, and r-IL-35 plus STAT4 inhibitors. The left image is a representative flow cytometry image, and the right image is a statistical graph ( n = 3 experiments). G RT-qPCR was used to detect the mRNA levels of NKG2A in NK-92MI cells treated with control, r-IL-35, r-IL-35 + STAT1 inhibitors, and rIL35 + STAT4 inhibitors ( n = 3 experiments). H Schematic diagram of STAT1 binding in the NKG2A promoter region. I ChIP was used to detect the binding of STAT1 to the NKG2A promoter region ( n = 3 experiments). J Schematic diagram of the molecular mechanism by which IL-35 upregulates NKG2A expression in NK cells via activating the STAT1 signaling pathway through its receptor ( A - D , F - H , J ) data are expressed as mean ± SD; B , F , J A two-tailed unpaired t-test was used; A , C , D , G , H A two-tailed unpaired t-test or one-way analysis of variance (ANOVA) and Tukey’s post hoc test was used

    Article Snippet: IKK inhibitor (BAY11-7082) (T1902), cGAS inhibitor (RU.521) (T5486), STAT1 inhibitor (Fludarabine) (T1038), and STAT4 inhibitor ((R)-Lisofylline)( T21694 ) were purchased from TargetMol, China.

    Techniques: Cell Culture, Quantitative RT-PCR, Flow Cytometry, Expressing, Control, Binding Assay, Two Tailed Test