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sting agonist addition  (MedChemExpress)


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    MedChemExpress sting agonist addition
    <t>STING</t> activation mainly drives autophagosome-independent LC3-lipidation that requires ATG16L1 lysine 490. LC3 lipidation (LC3-II FITC MFI) analyzed by flow cytometry ( A and B ) in splenocytes from WT or STING V154M/WT mutated mice ( A ) or tumor infiltrating CD4 + T cells from Fip200 fl/fl :cd4 Cre/+ mice, treated or not with i.t. injection of DMX (250 µg) for 5 h ( B ), n= 5-8 mice from 3 independent experiments, P values (*p<0.05, **p<0.01) determined by unpaired t-test. LC3 lipidation analyzed by western blot ( C-F ) in Fip200 + or ΔFip200 CD4 + T cells ( C ) stimulated or not ex vivo with DMXAA or 2’3’-cGAMP (DMX or cG) for 3 or 20 h, representative of n=2-3 independent experiments; in WT or Atg13 KO RAW264.7 macrophages ( D ) stimulated or not with DMX or cG for 3 or 20 h, representative of n=2-3 independent experiments; in adherent/myeloid cells isolated from human PBMCs ( E ) stimulated or not with cG for 3 h with or without <t>PI3KC3/VPS34</t> <t>inhibitor</t> (PI3KC3-IN1) or Bafilomycin A1, representative of n=3 donors; in WT or ATG16L1 K490A DC2.4 cells ( F ), stimulated or not with cG for 3, 6 or 20 h, representative of n=3 independent experiments. LC3 lipidation analyzed by immunofluorescence in WT or ATG16L1 K490A DC2.4 cells ( G ) stimulated or not with cG for 3 h, quantification of LC3 dots from ten fields of view, each containing >200 cells, **** P < 0.0001, and representative images, scale bar, 2 µm. LC3 lipidation analyzed by western blot ( H and I ) in WT or ATG16L1 K490A BMDCs ( H ) or CD4 + T cells ( I ), stimulated or not with cG for 3, 6 or 20 h, representative of 2 to 6 independent experiments. LC3-II:LC3-I ratios are indicated below western blot images and increases in LC3-II:LC3-I ratios, compared to control conditions, are highlighted in bold.
    Sting Agonist Addition, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sting agonist addition/product/MedChemExpress
    Average 94 stars, based on 11 article reviews
    sting agonist addition - by Bioz Stars, 2026-03
    94/100 stars

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    1) Product Images from "CASM potentiates STING-driven NFκB signaling in immune cells"

    Article Title: CASM potentiates STING-driven NFκB signaling in immune cells

    Journal: bioRxiv

    doi: 10.64898/2026.01.21.700774

    STING activation mainly drives autophagosome-independent LC3-lipidation that requires ATG16L1 lysine 490. LC3 lipidation (LC3-II FITC MFI) analyzed by flow cytometry ( A and B ) in splenocytes from WT or STING V154M/WT mutated mice ( A ) or tumor infiltrating CD4 + T cells from Fip200 fl/fl :cd4 Cre/+ mice, treated or not with i.t. injection of DMX (250 µg) for 5 h ( B ), n= 5-8 mice from 3 independent experiments, P values (*p<0.05, **p<0.01) determined by unpaired t-test. LC3 lipidation analyzed by western blot ( C-F ) in Fip200 + or ΔFip200 CD4 + T cells ( C ) stimulated or not ex vivo with DMXAA or 2’3’-cGAMP (DMX or cG) for 3 or 20 h, representative of n=2-3 independent experiments; in WT or Atg13 KO RAW264.7 macrophages ( D ) stimulated or not with DMX or cG for 3 or 20 h, representative of n=2-3 independent experiments; in adherent/myeloid cells isolated from human PBMCs ( E ) stimulated or not with cG for 3 h with or without PI3KC3/VPS34 inhibitor (PI3KC3-IN1) or Bafilomycin A1, representative of n=3 donors; in WT or ATG16L1 K490A DC2.4 cells ( F ), stimulated or not with cG for 3, 6 or 20 h, representative of n=3 independent experiments. LC3 lipidation analyzed by immunofluorescence in WT or ATG16L1 K490A DC2.4 cells ( G ) stimulated or not with cG for 3 h, quantification of LC3 dots from ten fields of view, each containing >200 cells, **** P < 0.0001, and representative images, scale bar, 2 µm. LC3 lipidation analyzed by western blot ( H and I ) in WT or ATG16L1 K490A BMDCs ( H ) or CD4 + T cells ( I ), stimulated or not with cG for 3, 6 or 20 h, representative of 2 to 6 independent experiments. LC3-II:LC3-I ratios are indicated below western blot images and increases in LC3-II:LC3-I ratios, compared to control conditions, are highlighted in bold.
    Figure Legend Snippet: STING activation mainly drives autophagosome-independent LC3-lipidation that requires ATG16L1 lysine 490. LC3 lipidation (LC3-II FITC MFI) analyzed by flow cytometry ( A and B ) in splenocytes from WT or STING V154M/WT mutated mice ( A ) or tumor infiltrating CD4 + T cells from Fip200 fl/fl :cd4 Cre/+ mice, treated or not with i.t. injection of DMX (250 µg) for 5 h ( B ), n= 5-8 mice from 3 independent experiments, P values (*p<0.05, **p<0.01) determined by unpaired t-test. LC3 lipidation analyzed by western blot ( C-F ) in Fip200 + or ΔFip200 CD4 + T cells ( C ) stimulated or not ex vivo with DMXAA or 2’3’-cGAMP (DMX or cG) for 3 or 20 h, representative of n=2-3 independent experiments; in WT or Atg13 KO RAW264.7 macrophages ( D ) stimulated or not with DMX or cG for 3 or 20 h, representative of n=2-3 independent experiments; in adherent/myeloid cells isolated from human PBMCs ( E ) stimulated or not with cG for 3 h with or without PI3KC3/VPS34 inhibitor (PI3KC3-IN1) or Bafilomycin A1, representative of n=3 donors; in WT or ATG16L1 K490A DC2.4 cells ( F ), stimulated or not with cG for 3, 6 or 20 h, representative of n=3 independent experiments. LC3 lipidation analyzed by immunofluorescence in WT or ATG16L1 K490A DC2.4 cells ( G ) stimulated or not with cG for 3 h, quantification of LC3 dots from ten fields of view, each containing >200 cells, **** P < 0.0001, and representative images, scale bar, 2 µm. LC3 lipidation analyzed by western blot ( H and I ) in WT or ATG16L1 K490A BMDCs ( H ) or CD4 + T cells ( I ), stimulated or not with cG for 3, 6 or 20 h, representative of 2 to 6 independent experiments. LC3-II:LC3-I ratios are indicated below western blot images and increases in LC3-II:LC3-I ratios, compared to control conditions, are highlighted in bold.

    Techniques Used: Activation Assay, Flow Cytometry, Injection, Western Blot, Ex Vivo, Isolation, Immunofluorescence, Control

    STING-driven CASM is induced by various STING ligands and requires STING but not IRF3 or RUBICON. LC3 lipidation analyzed by western blot ( A to D) in WT or Sting KO BMDCs ( A ) or CD4 + T cells ( B ), stimulated or not with DMX, cG, cdi-AMP or cdi-GMP for 3 h, representative of n=2 to 5 independent experiments; in WT or Irf3 KO BMDCs ( C ) or CD4 + T cells ( D ), stimulated or not with cG for 3 h, representative of n=3 independent experiments. LC3 lipidation (LC3-FITC MFI, normalized to Bafilomycin A1 condition) analyzed by flow cytometry ( E ) in WT or Rubcn KO CD11b + cells, CD4 + or CD8 + T cells, stimulated or not with DMX for 3 h, n= 5 mice from 3 independent experiments, P values (*p<0.05, **p<0.01) determined by two-way ANOVA. LC3-II:LC3-I ratios are indicated below western blot images and increases in LC3-II:LC3-I ratios, compared to control conditions, are highlighted in bold.
    Figure Legend Snippet: STING-driven CASM is induced by various STING ligands and requires STING but not IRF3 or RUBICON. LC3 lipidation analyzed by western blot ( A to D) in WT or Sting KO BMDCs ( A ) or CD4 + T cells ( B ), stimulated or not with DMX, cG, cdi-AMP or cdi-GMP for 3 h, representative of n=2 to 5 independent experiments; in WT or Irf3 KO BMDCs ( C ) or CD4 + T cells ( D ), stimulated or not with cG for 3 h, representative of n=3 independent experiments. LC3 lipidation (LC3-FITC MFI, normalized to Bafilomycin A1 condition) analyzed by flow cytometry ( E ) in WT or Rubcn KO CD11b + cells, CD4 + or CD8 + T cells, stimulated or not with DMX for 3 h, n= 5 mice from 3 independent experiments, P values (*p<0.05, **p<0.01) determined by two-way ANOVA. LC3-II:LC3-I ratios are indicated below western blot images and increases in LC3-II:LC3-I ratios, compared to control conditions, are highlighted in bold.

    Techniques Used: Western Blot, Flow Cytometry, Control

    STING-associated immune responses are upregulated in autophagy-incompetent macrophages and T cells. ( A ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from WT and Atg13 KO RAW264.7 macrophages stimulated or not with cG for 3 h. ( B ) IFN-β and CXCL10 secretion measured by ELISA from WT and Atg13 KO RAW264.7 macrophages stimulated or not with cG for 3 h or 6 h respectively. ( C ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from Fip200 + and ΔFip200 CD4 + T cells stimulated or not with cG for 20 h. ( A and C ) Pooled data from 4 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA from each target relative expression. ( D ) IFN-β, CXCL10 and IL-6 secretion measured by ELISA from Fip200 + and ΔFip200 CD4 + T cells stimulated or not with cG 20 h. ( B and D ), pooled data (Mean +/- SD) from 2 to 5 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA. ( E and F ) Western blot analyses of indicated proteins and corresponding relevant quantifications from WT and Atg13 KO RAW264.7 macrophages ( E ) or Fip200 + and ΔFip200 CD4 + T cells ( F ) stimulated or not with cG for indicated time, pooled data (Mean +/- SEM) from 3 to 5 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA.
    Figure Legend Snippet: STING-associated immune responses are upregulated in autophagy-incompetent macrophages and T cells. ( A ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from WT and Atg13 KO RAW264.7 macrophages stimulated or not with cG for 3 h. ( B ) IFN-β and CXCL10 secretion measured by ELISA from WT and Atg13 KO RAW264.7 macrophages stimulated or not with cG for 3 h or 6 h respectively. ( C ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from Fip200 + and ΔFip200 CD4 + T cells stimulated or not with cG for 20 h. ( A and C ) Pooled data from 4 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA from each target relative expression. ( D ) IFN-β, CXCL10 and IL-6 secretion measured by ELISA from Fip200 + and ΔFip200 CD4 + T cells stimulated or not with cG 20 h. ( B and D ), pooled data (Mean +/- SD) from 2 to 5 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA. ( E and F ) Western blot analyses of indicated proteins and corresponding relevant quantifications from WT and Atg13 KO RAW264.7 macrophages ( E ) or Fip200 + and ΔFip200 CD4 + T cells ( F ) stimulated or not with cG for indicated time, pooled data (Mean +/- SEM) from 3 to 5 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA.

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

    STING-driven CASM regulates RELA/p65-induced inflammatory response in myeloid cells. ( A ) GSEA (Gene set enrichment analysis; Normalized Enrichment Score (NES) & -log10 (p adj )) obtained from RNA sequencing analysis of WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 h. ( B ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 or 20 h, pooled data (Mean) from n=6-7 mice from 3 independent experiments, P values (*p<0.05) determined by two-way ANOVA from each target relative expression. IFN-β secretion measured by ELISA ( C ) or western blot analysis of indicated proteins and corresponding quantifications ( D ) from WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 ( D ) or 20 h ( C and D ), pooled data (Mean +/- SD) from n=5 mice from 2 independent experiments, P values (**p<0.01, ***p<0.001, ****p<0.0001) determined by two-way ANOVA. Tnfa mRNA expression determined by RTqPCR (Fold Change FC; Normalized to mean of each control condition) ( E ) and TNF-α secretion measured by ELISA ( F ) from WT or ATG16L1 K490A DC2.4 cells stimulated or not with cG for 6 h, pooled data (Mean +/- SD) from 3 to 5 independent experiments, P values (*p<0.05, ***p<0.001, ****p<0.0001) determined by two-way ANOVA. ( G ) Heatmap representing indicated molecule secretion (Z-score) measured by Cytokine Array from WT or ATG16L1 K490A DC2.4 cells stimulated or not with cG for 6 h, pooled data from 2 independent experiments. Western blot analysis of indicated proteins from total ( H and I ) or nuclear ( J ) extracts and corresponding quantifications (Fold Change FC, normalized to mean of WT mouse control condition) from WT or ATG16L1 K490A DC2.4 cells ( H and J ) or BMDCs ( I ) stimulated or not with cG for 3 h, pooled data (Mean & SD) from 5 independent experiments ( H ) or from n=3 mice ( I and J ), P values (*p<0.05, **p<0.01) determined by two-way ANOVA.
    Figure Legend Snippet: STING-driven CASM regulates RELA/p65-induced inflammatory response in myeloid cells. ( A ) GSEA (Gene set enrichment analysis; Normalized Enrichment Score (NES) & -log10 (p adj )) obtained from RNA sequencing analysis of WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 h. ( B ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 or 20 h, pooled data (Mean) from n=6-7 mice from 3 independent experiments, P values (*p<0.05) determined by two-way ANOVA from each target relative expression. IFN-β secretion measured by ELISA ( C ) or western blot analysis of indicated proteins and corresponding quantifications ( D ) from WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 ( D ) or 20 h ( C and D ), pooled data (Mean +/- SD) from n=5 mice from 2 independent experiments, P values (**p<0.01, ***p<0.001, ****p<0.0001) determined by two-way ANOVA. Tnfa mRNA expression determined by RTqPCR (Fold Change FC; Normalized to mean of each control condition) ( E ) and TNF-α secretion measured by ELISA ( F ) from WT or ATG16L1 K490A DC2.4 cells stimulated or not with cG for 6 h, pooled data (Mean +/- SD) from 3 to 5 independent experiments, P values (*p<0.05, ***p<0.001, ****p<0.0001) determined by two-way ANOVA. ( G ) Heatmap representing indicated molecule secretion (Z-score) measured by Cytokine Array from WT or ATG16L1 K490A DC2.4 cells stimulated or not with cG for 6 h, pooled data from 2 independent experiments. Western blot analysis of indicated proteins from total ( H and I ) or nuclear ( J ) extracts and corresponding quantifications (Fold Change FC, normalized to mean of WT mouse control condition) from WT or ATG16L1 K490A DC2.4 cells ( H and J ) or BMDCs ( I ) stimulated or not with cG for 3 h, pooled data (Mean & SD) from 5 independent experiments ( H ) or from n=3 mice ( I and J ), P values (*p<0.05, **p<0.01) determined by two-way ANOVA.

    Techniques Used: RNA Sequencing, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Control



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    sting agonist addition  (MedChemExpress)
    94
    MedChemExpress sting agonist addition
    <t>STING</t> activation mainly drives autophagosome-independent LC3-lipidation that requires ATG16L1 lysine 490. LC3 lipidation (LC3-II FITC MFI) analyzed by flow cytometry ( A and B ) in splenocytes from WT or STING V154M/WT mutated mice ( A ) or tumor infiltrating CD4 + T cells from Fip200 fl/fl :cd4 Cre/+ mice, treated or not with i.t. injection of DMX (250 µg) for 5 h ( B ), n= 5-8 mice from 3 independent experiments, P values (*p<0.05, **p<0.01) determined by unpaired t-test. LC3 lipidation analyzed by western blot ( C-F ) in Fip200 + or ΔFip200 CD4 + T cells ( C ) stimulated or not ex vivo with DMXAA or 2’3’-cGAMP (DMX or cG) for 3 or 20 h, representative of n=2-3 independent experiments; in WT or Atg13 KO RAW264.7 macrophages ( D ) stimulated or not with DMX or cG for 3 or 20 h, representative of n=2-3 independent experiments; in adherent/myeloid cells isolated from human PBMCs ( E ) stimulated or not with cG for 3 h with or without <t>PI3KC3/VPS34</t> <t>inhibitor</t> (PI3KC3-IN1) or Bafilomycin A1, representative of n=3 donors; in WT or ATG16L1 K490A DC2.4 cells ( F ), stimulated or not with cG for 3, 6 or 20 h, representative of n=3 independent experiments. LC3 lipidation analyzed by immunofluorescence in WT or ATG16L1 K490A DC2.4 cells ( G ) stimulated or not with cG for 3 h, quantification of LC3 dots from ten fields of view, each containing >200 cells, **** P < 0.0001, and representative images, scale bar, 2 µm. LC3 lipidation analyzed by western blot ( H and I ) in WT or ATG16L1 K490A BMDCs ( H ) or CD4 + T cells ( I ), stimulated or not with cG for 3, 6 or 20 h, representative of 2 to 6 independent experiments. LC3-II:LC3-I ratios are indicated below western blot images and increases in LC3-II:LC3-I ratios, compared to control conditions, are highlighted in bold.
    Sting Agonist Addition, supplied by MedChemExpress, 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/result/sting agonist addition/product/MedChemExpress
    Average 94 stars, based on 1 article reviews
    sting agonist addition - by Bioz Stars, 2026-03
    94/100 stars
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    STING activation mainly drives autophagosome-independent LC3-lipidation that requires ATG16L1 lysine 490. LC3 lipidation (LC3-II FITC MFI) analyzed by flow cytometry ( A and B ) in splenocytes from WT or STING V154M/WT mutated mice ( A ) or tumor infiltrating CD4 + T cells from Fip200 fl/fl :cd4 Cre/+ mice, treated or not with i.t. injection of DMX (250 µg) for 5 h ( B ), n= 5-8 mice from 3 independent experiments, P values (*p<0.05, **p<0.01) determined by unpaired t-test. LC3 lipidation analyzed by western blot ( C-F ) in Fip200 + or ΔFip200 CD4 + T cells ( C ) stimulated or not ex vivo with DMXAA or 2’3’-cGAMP (DMX or cG) for 3 or 20 h, representative of n=2-3 independent experiments; in WT or Atg13 KO RAW264.7 macrophages ( D ) stimulated or not with DMX or cG for 3 or 20 h, representative of n=2-3 independent experiments; in adherent/myeloid cells isolated from human PBMCs ( E ) stimulated or not with cG for 3 h with or without PI3KC3/VPS34 inhibitor (PI3KC3-IN1) or Bafilomycin A1, representative of n=3 donors; in WT or ATG16L1 K490A DC2.4 cells ( F ), stimulated or not with cG for 3, 6 or 20 h, representative of n=3 independent experiments. LC3 lipidation analyzed by immunofluorescence in WT or ATG16L1 K490A DC2.4 cells ( G ) stimulated or not with cG for 3 h, quantification of LC3 dots from ten fields of view, each containing >200 cells, **** P < 0.0001, and representative images, scale bar, 2 µm. LC3 lipidation analyzed by western blot ( H and I ) in WT or ATG16L1 K490A BMDCs ( H ) or CD4 + T cells ( I ), stimulated or not with cG for 3, 6 or 20 h, representative of 2 to 6 independent experiments. LC3-II:LC3-I ratios are indicated below western blot images and increases in LC3-II:LC3-I ratios, compared to control conditions, are highlighted in bold.

    Journal: bioRxiv

    Article Title: CASM potentiates STING-driven NFκB signaling in immune cells

    doi: 10.64898/2026.01.21.700774

    Figure Lengend Snippet: STING activation mainly drives autophagosome-independent LC3-lipidation that requires ATG16L1 lysine 490. LC3 lipidation (LC3-II FITC MFI) analyzed by flow cytometry ( A and B ) in splenocytes from WT or STING V154M/WT mutated mice ( A ) or tumor infiltrating CD4 + T cells from Fip200 fl/fl :cd4 Cre/+ mice, treated or not with i.t. injection of DMX (250 µg) for 5 h ( B ), n= 5-8 mice from 3 independent experiments, P values (*p<0.05, **p<0.01) determined by unpaired t-test. LC3 lipidation analyzed by western blot ( C-F ) in Fip200 + or ΔFip200 CD4 + T cells ( C ) stimulated or not ex vivo with DMXAA or 2’3’-cGAMP (DMX or cG) for 3 or 20 h, representative of n=2-3 independent experiments; in WT or Atg13 KO RAW264.7 macrophages ( D ) stimulated or not with DMX or cG for 3 or 20 h, representative of n=2-3 independent experiments; in adherent/myeloid cells isolated from human PBMCs ( E ) stimulated or not with cG for 3 h with or without PI3KC3/VPS34 inhibitor (PI3KC3-IN1) or Bafilomycin A1, representative of n=3 donors; in WT or ATG16L1 K490A DC2.4 cells ( F ), stimulated or not with cG for 3, 6 or 20 h, representative of n=3 independent experiments. LC3 lipidation analyzed by immunofluorescence in WT or ATG16L1 K490A DC2.4 cells ( G ) stimulated or not with cG for 3 h, quantification of LC3 dots from ten fields of view, each containing >200 cells, **** P < 0.0001, and representative images, scale bar, 2 µm. LC3 lipidation analyzed by western blot ( H and I ) in WT or ATG16L1 K490A BMDCs ( H ) or CD4 + T cells ( I ), stimulated or not with cG for 3, 6 or 20 h, representative of 2 to 6 independent experiments. LC3-II:LC3-I ratios are indicated below western blot images and increases in LC3-II:LC3-I ratios, compared to control conditions, are highlighted in bold.

    Article Snippet: TBK1/IKKε inhibition was performed by pre-treating cells overnight with 0.5 μM of BX795 (TBK1/IKKε inhibitor - InvitroFitTM Invivogen, tlr-bx7) before STING agonist addition for 3 h. To distinguish between autophagy and CASM, cells were pre-treated with Bafilomycin A1 (BafA1, 100 nM, Tocris, 1334) or PIK3C3-IN1 (1 μM, MedChemExpress, HY-12795) for 1 h and co-treated with cG for 3 additional hours.

    Techniques: Activation Assay, Flow Cytometry, Injection, Western Blot, Ex Vivo, Isolation, Immunofluorescence, Control

    STING-driven CASM is induced by various STING ligands and requires STING but not IRF3 or RUBICON. LC3 lipidation analyzed by western blot ( A to D) in WT or Sting KO BMDCs ( A ) or CD4 + T cells ( B ), stimulated or not with DMX, cG, cdi-AMP or cdi-GMP for 3 h, representative of n=2 to 5 independent experiments; in WT or Irf3 KO BMDCs ( C ) or CD4 + T cells ( D ), stimulated or not with cG for 3 h, representative of n=3 independent experiments. LC3 lipidation (LC3-FITC MFI, normalized to Bafilomycin A1 condition) analyzed by flow cytometry ( E ) in WT or Rubcn KO CD11b + cells, CD4 + or CD8 + T cells, stimulated or not with DMX for 3 h, n= 5 mice from 3 independent experiments, P values (*p<0.05, **p<0.01) determined by two-way ANOVA. LC3-II:LC3-I ratios are indicated below western blot images and increases in LC3-II:LC3-I ratios, compared to control conditions, are highlighted in bold.

    Journal: bioRxiv

    Article Title: CASM potentiates STING-driven NFκB signaling in immune cells

    doi: 10.64898/2026.01.21.700774

    Figure Lengend Snippet: STING-driven CASM is induced by various STING ligands and requires STING but not IRF3 or RUBICON. LC3 lipidation analyzed by western blot ( A to D) in WT or Sting KO BMDCs ( A ) or CD4 + T cells ( B ), stimulated or not with DMX, cG, cdi-AMP or cdi-GMP for 3 h, representative of n=2 to 5 independent experiments; in WT or Irf3 KO BMDCs ( C ) or CD4 + T cells ( D ), stimulated or not with cG for 3 h, representative of n=3 independent experiments. LC3 lipidation (LC3-FITC MFI, normalized to Bafilomycin A1 condition) analyzed by flow cytometry ( E ) in WT or Rubcn KO CD11b + cells, CD4 + or CD8 + T cells, stimulated or not with DMX for 3 h, n= 5 mice from 3 independent experiments, P values (*p<0.05, **p<0.01) determined by two-way ANOVA. LC3-II:LC3-I ratios are indicated below western blot images and increases in LC3-II:LC3-I ratios, compared to control conditions, are highlighted in bold.

    Article Snippet: TBK1/IKKε inhibition was performed by pre-treating cells overnight with 0.5 μM of BX795 (TBK1/IKKε inhibitor - InvitroFitTM Invivogen, tlr-bx7) before STING agonist addition for 3 h. To distinguish between autophagy and CASM, cells were pre-treated with Bafilomycin A1 (BafA1, 100 nM, Tocris, 1334) or PIK3C3-IN1 (1 μM, MedChemExpress, HY-12795) for 1 h and co-treated with cG for 3 additional hours.

    Techniques: Western Blot, Flow Cytometry, Control

    STING-associated immune responses are upregulated in autophagy-incompetent macrophages and T cells. ( A ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from WT and Atg13 KO RAW264.7 macrophages stimulated or not with cG for 3 h. ( B ) IFN-β and CXCL10 secretion measured by ELISA from WT and Atg13 KO RAW264.7 macrophages stimulated or not with cG for 3 h or 6 h respectively. ( C ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from Fip200 + and ΔFip200 CD4 + T cells stimulated or not with cG for 20 h. ( A and C ) Pooled data from 4 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA from each target relative expression. ( D ) IFN-β, CXCL10 and IL-6 secretion measured by ELISA from Fip200 + and ΔFip200 CD4 + T cells stimulated or not with cG 20 h. ( B and D ), pooled data (Mean +/- SD) from 2 to 5 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA. ( E and F ) Western blot analyses of indicated proteins and corresponding relevant quantifications from WT and Atg13 KO RAW264.7 macrophages ( E ) or Fip200 + and ΔFip200 CD4 + T cells ( F ) stimulated or not with cG for indicated time, pooled data (Mean +/- SEM) from 3 to 5 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA.

    Journal: bioRxiv

    Article Title: CASM potentiates STING-driven NFκB signaling in immune cells

    doi: 10.64898/2026.01.21.700774

    Figure Lengend Snippet: STING-associated immune responses are upregulated in autophagy-incompetent macrophages and T cells. ( A ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from WT and Atg13 KO RAW264.7 macrophages stimulated or not with cG for 3 h. ( B ) IFN-β and CXCL10 secretion measured by ELISA from WT and Atg13 KO RAW264.7 macrophages stimulated or not with cG for 3 h or 6 h respectively. ( C ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from Fip200 + and ΔFip200 CD4 + T cells stimulated or not with cG for 20 h. ( A and C ) Pooled data from 4 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA from each target relative expression. ( D ) IFN-β, CXCL10 and IL-6 secretion measured by ELISA from Fip200 + and ΔFip200 CD4 + T cells stimulated or not with cG 20 h. ( B and D ), pooled data (Mean +/- SD) from 2 to 5 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA. ( E and F ) Western blot analyses of indicated proteins and corresponding relevant quantifications from WT and Atg13 KO RAW264.7 macrophages ( E ) or Fip200 + and ΔFip200 CD4 + T cells ( F ) stimulated or not with cG for indicated time, pooled data (Mean +/- SEM) from 3 to 5 independent experiments, P values (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not represented) determined by two-way ANOVA.

    Article Snippet: TBK1/IKKε inhibition was performed by pre-treating cells overnight with 0.5 μM of BX795 (TBK1/IKKε inhibitor - InvitroFitTM Invivogen, tlr-bx7) before STING agonist addition for 3 h. To distinguish between autophagy and CASM, cells were pre-treated with Bafilomycin A1 (BafA1, 100 nM, Tocris, 1334) or PIK3C3-IN1 (1 μM, MedChemExpress, HY-12795) for 1 h and co-treated with cG for 3 additional hours.

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

    STING-driven CASM regulates RELA/p65-induced inflammatory response in myeloid cells. ( A ) GSEA (Gene set enrichment analysis; Normalized Enrichment Score (NES) & -log10 (p adj )) obtained from RNA sequencing analysis of WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 h. ( B ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 or 20 h, pooled data (Mean) from n=6-7 mice from 3 independent experiments, P values (*p<0.05) determined by two-way ANOVA from each target relative expression. IFN-β secretion measured by ELISA ( C ) or western blot analysis of indicated proteins and corresponding quantifications ( D ) from WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 ( D ) or 20 h ( C and D ), pooled data (Mean +/- SD) from n=5 mice from 2 independent experiments, P values (**p<0.01, ***p<0.001, ****p<0.0001) determined by two-way ANOVA. Tnfa mRNA expression determined by RTqPCR (Fold Change FC; Normalized to mean of each control condition) ( E ) and TNF-α secretion measured by ELISA ( F ) from WT or ATG16L1 K490A DC2.4 cells stimulated or not with cG for 6 h, pooled data (Mean +/- SD) from 3 to 5 independent experiments, P values (*p<0.05, ***p<0.001, ****p<0.0001) determined by two-way ANOVA. ( G ) Heatmap representing indicated molecule secretion (Z-score) measured by Cytokine Array from WT or ATG16L1 K490A DC2.4 cells stimulated or not with cG for 6 h, pooled data from 2 independent experiments. Western blot analysis of indicated proteins from total ( H and I ) or nuclear ( J ) extracts and corresponding quantifications (Fold Change FC, normalized to mean of WT mouse control condition) from WT or ATG16L1 K490A DC2.4 cells ( H and J ) or BMDCs ( I ) stimulated or not with cG for 3 h, pooled data (Mean & SD) from 5 independent experiments ( H ) or from n=3 mice ( I and J ), P values (*p<0.05, **p<0.01) determined by two-way ANOVA.

    Journal: bioRxiv

    Article Title: CASM potentiates STING-driven NFκB signaling in immune cells

    doi: 10.64898/2026.01.21.700774

    Figure Lengend Snippet: STING-driven CASM regulates RELA/p65-induced inflammatory response in myeloid cells. ( A ) GSEA (Gene set enrichment analysis; Normalized Enrichment Score (NES) & -log10 (p adj )) obtained from RNA sequencing analysis of WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 h. ( B ) Heatmap representing indicated cytokine and ISG mRNA expression (Z-score) determined by RTqPCR from WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 or 20 h, pooled data (Mean) from n=6-7 mice from 3 independent experiments, P values (*p<0.05) determined by two-way ANOVA from each target relative expression. IFN-β secretion measured by ELISA ( C ) or western blot analysis of indicated proteins and corresponding quantifications ( D ) from WT or ATG16L1 K490A BMDCs stimulated or not with cG for 3 ( D ) or 20 h ( C and D ), pooled data (Mean +/- SD) from n=5 mice from 2 independent experiments, P values (**p<0.01, ***p<0.001, ****p<0.0001) determined by two-way ANOVA. Tnfa mRNA expression determined by RTqPCR (Fold Change FC; Normalized to mean of each control condition) ( E ) and TNF-α secretion measured by ELISA ( F ) from WT or ATG16L1 K490A DC2.4 cells stimulated or not with cG for 6 h, pooled data (Mean +/- SD) from 3 to 5 independent experiments, P values (*p<0.05, ***p<0.001, ****p<0.0001) determined by two-way ANOVA. ( G ) Heatmap representing indicated molecule secretion (Z-score) measured by Cytokine Array from WT or ATG16L1 K490A DC2.4 cells stimulated or not with cG for 6 h, pooled data from 2 independent experiments. Western blot analysis of indicated proteins from total ( H and I ) or nuclear ( J ) extracts and corresponding quantifications (Fold Change FC, normalized to mean of WT mouse control condition) from WT or ATG16L1 K490A DC2.4 cells ( H and J ) or BMDCs ( I ) stimulated or not with cG for 3 h, pooled data (Mean & SD) from 5 independent experiments ( H ) or from n=3 mice ( I and J ), P values (*p<0.05, **p<0.01) determined by two-way ANOVA.

    Article Snippet: TBK1/IKKε inhibition was performed by pre-treating cells overnight with 0.5 μM of BX795 (TBK1/IKKε inhibitor - InvitroFitTM Invivogen, tlr-bx7) before STING agonist addition for 3 h. To distinguish between autophagy and CASM, cells were pre-treated with Bafilomycin A1 (BafA1, 100 nM, Tocris, 1334) or PIK3C3-IN1 (1 μM, MedChemExpress, HY-12795) for 1 h and co-treated with cG for 3 additional hours.

    Techniques: RNA Sequencing, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Control