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gfp signals  (Revvity)


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    Revvity gfp signals
    a Hypothesis diagram depicting the crosstalk between immune inflammatory cells and PDAC cells. b Heatmap of transcriptome DEGs in the cellular response to IFN-γ process between KPC-shScramble (Scr) cells and KPC-shUbe2d3-1 (KD1) cells ( n = 3 samples per group). c Tumor growth curves following <t>subcutaneous</t> <t>inoculation</t> of KPC/Panc02 Scr/KD1 cells in syngeneic immunodeficient Rag1 −/− mice and subsequent implantation into immunocompetent C57BL/6 mice. Once tumor reached 1000 mm 3 in Rag1 −/− mice ( n = 4 mice per group), they were excised, and 1–2 mm 3 fragments were further inoculated subcutaneously into immunocompetent C57BL/6 mice ( n = 7 mice per group). d Experimental setup for in vitro CTL-mediated killing of KPC-WT/KO cells. KPC-WT cells were subjected to lentiviral transduction with the indicated sgCtrl <t>(Ctrl)-GFP,</t> sgCtrl (Ctrl)-mCherry, sgUbe2d3-1 (KO1)-mCherry, or sgUbe2d3-2 (KO2)-mCherry Cas9 vectors. These cells were further transiently transfected with the OVA OE pcDNA3 plasmid to construct target cells. OT-I CD8 + T cells were cocultured with either a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -Ctrl-mCherry cells or a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -KO-mCherry cells for 24 h ( n = 5 independent experiments per group). e Quantification of tumor-infiltrating CD4 + T cells and CD8 + T cells in subcutaneous KPC Scr/KD1 model ( n = 5 mice per group) and Panc02 Scr/KD1 model ( n = 9 mice per group). f Tumor growth curves from CD4 + or CD8 + T-cell deletion assay ( n = 5 mice per group). g Tumor growth evaluation in humanized NCG mice. Seven days post-PBMC injection, these immunoreconstituted mice were subcutaneously engrafted with Capan-1-Scr or Capan-1-KD1 cells. The level of immune humanization was confirmed by flow cytometric analysis of hCD45 + cells in peripheral blood on days 7 and 21. All mice were sacrificed 24 days after tumor inoculation for tumor weight measurement ( n = 7 mice per group). Statistical significance was determined via the unpaired t test (two-tailed) in ( c – g ). The data are presented as the means ± SDs. Source data are provided in the Source Data file.
    Gfp Signals, supplied by Revvity, used in various techniques. Bioz Stars score: 99/100, based on 37484 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "IFN-γ-driven UBE2D3 upregulation impairs antigen presentation pathways and anti-tumor immunity in pancreatic cancer"

    Article Title: IFN-γ-driven UBE2D3 upregulation impairs antigen presentation pathways and anti-tumor immunity in pancreatic cancer

    Journal: Nature Communications

    doi: 10.1038/s41467-025-65762-4

    a Hypothesis diagram depicting the crosstalk between immune inflammatory cells and PDAC cells. b Heatmap of transcriptome DEGs in the cellular response to IFN-γ process between KPC-shScramble (Scr) cells and KPC-shUbe2d3-1 (KD1) cells ( n = 3 samples per group). c Tumor growth curves following subcutaneous inoculation of KPC/Panc02 Scr/KD1 cells in syngeneic immunodeficient Rag1 −/− mice and subsequent implantation into immunocompetent C57BL/6 mice. Once tumor reached 1000 mm 3 in Rag1 −/− mice ( n = 4 mice per group), they were excised, and 1–2 mm 3 fragments were further inoculated subcutaneously into immunocompetent C57BL/6 mice ( n = 7 mice per group). d Experimental setup for in vitro CTL-mediated killing of KPC-WT/KO cells. KPC-WT cells were subjected to lentiviral transduction with the indicated sgCtrl (Ctrl)-GFP, sgCtrl (Ctrl)-mCherry, sgUbe2d3-1 (KO1)-mCherry, or sgUbe2d3-2 (KO2)-mCherry Cas9 vectors. These cells were further transiently transfected with the OVA OE pcDNA3 plasmid to construct target cells. OT-I CD8 + T cells were cocultured with either a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -Ctrl-mCherry cells or a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -KO-mCherry cells for 24 h ( n = 5 independent experiments per group). e Quantification of tumor-infiltrating CD4 + T cells and CD8 + T cells in subcutaneous KPC Scr/KD1 model ( n = 5 mice per group) and Panc02 Scr/KD1 model ( n = 9 mice per group). f Tumor growth curves from CD4 + or CD8 + T-cell deletion assay ( n = 5 mice per group). g Tumor growth evaluation in humanized NCG mice. Seven days post-PBMC injection, these immunoreconstituted mice were subcutaneously engrafted with Capan-1-Scr or Capan-1-KD1 cells. The level of immune humanization was confirmed by flow cytometric analysis of hCD45 + cells in peripheral blood on days 7 and 21. All mice were sacrificed 24 days after tumor inoculation for tumor weight measurement ( n = 7 mice per group). Statistical significance was determined via the unpaired t test (two-tailed) in ( c – g ). The data are presented as the means ± SDs. Source data are provided in the Source Data file.
    Figure Legend Snippet: a Hypothesis diagram depicting the crosstalk between immune inflammatory cells and PDAC cells. b Heatmap of transcriptome DEGs in the cellular response to IFN-γ process between KPC-shScramble (Scr) cells and KPC-shUbe2d3-1 (KD1) cells ( n = 3 samples per group). c Tumor growth curves following subcutaneous inoculation of KPC/Panc02 Scr/KD1 cells in syngeneic immunodeficient Rag1 −/− mice and subsequent implantation into immunocompetent C57BL/6 mice. Once tumor reached 1000 mm 3 in Rag1 −/− mice ( n = 4 mice per group), they were excised, and 1–2 mm 3 fragments were further inoculated subcutaneously into immunocompetent C57BL/6 mice ( n = 7 mice per group). d Experimental setup for in vitro CTL-mediated killing of KPC-WT/KO cells. KPC-WT cells were subjected to lentiviral transduction with the indicated sgCtrl (Ctrl)-GFP, sgCtrl (Ctrl)-mCherry, sgUbe2d3-1 (KO1)-mCherry, or sgUbe2d3-2 (KO2)-mCherry Cas9 vectors. These cells were further transiently transfected with the OVA OE pcDNA3 plasmid to construct target cells. OT-I CD8 + T cells were cocultured with either a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -Ctrl-mCherry cells or a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -KO-mCherry cells for 24 h ( n = 5 independent experiments per group). e Quantification of tumor-infiltrating CD4 + T cells and CD8 + T cells in subcutaneous KPC Scr/KD1 model ( n = 5 mice per group) and Panc02 Scr/KD1 model ( n = 9 mice per group). f Tumor growth curves from CD4 + or CD8 + T-cell deletion assay ( n = 5 mice per group). g Tumor growth evaluation in humanized NCG mice. Seven days post-PBMC injection, these immunoreconstituted mice were subcutaneously engrafted with Capan-1-Scr or Capan-1-KD1 cells. The level of immune humanization was confirmed by flow cytometric analysis of hCD45 + cells in peripheral blood on days 7 and 21. All mice were sacrificed 24 days after tumor inoculation for tumor weight measurement ( n = 7 mice per group). Statistical significance was determined via the unpaired t test (two-tailed) in ( c – g ). The data are presented as the means ± SDs. Source data are provided in the Source Data file.

    Techniques Used: In Vitro, Transduction, Transfection, Plasmid Preparation, Construct, DNA Deletion Assay, Injection, Two Tailed Test



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    a Hypothesis diagram depicting the crosstalk between immune inflammatory cells and PDAC cells. b Heatmap of transcriptome DEGs in the cellular response to IFN-γ process between KPC-shScramble (Scr) cells and KPC-shUbe2d3-1 (KD1) cells ( n = 3 samples per group). c Tumor growth curves following <t>subcutaneous</t> <t>inoculation</t> of KPC/Panc02 Scr/KD1 cells in syngeneic immunodeficient Rag1 −/− mice and subsequent implantation into immunocompetent C57BL/6 mice. Once tumor reached 1000 mm 3 in Rag1 −/− mice ( n = 4 mice per group), they were excised, and 1–2 mm 3 fragments were further inoculated subcutaneously into immunocompetent C57BL/6 mice ( n = 7 mice per group). d Experimental setup for in vitro CTL-mediated killing of KPC-WT/KO cells. KPC-WT cells were subjected to lentiviral transduction with the indicated sgCtrl <t>(Ctrl)-GFP,</t> sgCtrl (Ctrl)-mCherry, sgUbe2d3-1 (KO1)-mCherry, or sgUbe2d3-2 (KO2)-mCherry Cas9 vectors. These cells were further transiently transfected with the OVA OE pcDNA3 plasmid to construct target cells. OT-I CD8 + T cells were cocultured with either a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -Ctrl-mCherry cells or a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -KO-mCherry cells for 24 h ( n = 5 independent experiments per group). e Quantification of tumor-infiltrating CD4 + T cells and CD8 + T cells in subcutaneous KPC Scr/KD1 model ( n = 5 mice per group) and Panc02 Scr/KD1 model ( n = 9 mice per group). f Tumor growth curves from CD4 + or CD8 + T-cell deletion assay ( n = 5 mice per group). g Tumor growth evaluation in humanized NCG mice. Seven days post-PBMC injection, these immunoreconstituted mice were subcutaneously engrafted with Capan-1-Scr or Capan-1-KD1 cells. The level of immune humanization was confirmed by flow cytometric analysis of hCD45 + cells in peripheral blood on days 7 and 21. All mice were sacrificed 24 days after tumor inoculation for tumor weight measurement ( n = 7 mice per group). Statistical significance was determined via the unpaired t test (two-tailed) in ( c – g ). The data are presented as the means ± SDs. Source data are provided in the Source Data file.
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    a Hypothesis diagram depicting the crosstalk between immune inflammatory cells and PDAC cells. b Heatmap of transcriptome DEGs in the cellular response to IFN-γ process between KPC-shScramble (Scr) cells and KPC-shUbe2d3-1 (KD1) cells ( n = 3 samples per group). c Tumor growth curves following <t>subcutaneous</t> <t>inoculation</t> of KPC/Panc02 Scr/KD1 cells in syngeneic immunodeficient Rag1 −/− mice and subsequent implantation into immunocompetent C57BL/6 mice. Once tumor reached 1000 mm 3 in Rag1 −/− mice ( n = 4 mice per group), they were excised, and 1–2 mm 3 fragments were further inoculated subcutaneously into immunocompetent C57BL/6 mice ( n = 7 mice per group). d Experimental setup for in vitro CTL-mediated killing of KPC-WT/KO cells. KPC-WT cells were subjected to lentiviral transduction with the indicated sgCtrl <t>(Ctrl)-GFP,</t> sgCtrl (Ctrl)-mCherry, sgUbe2d3-1 (KO1)-mCherry, or sgUbe2d3-2 (KO2)-mCherry Cas9 vectors. These cells were further transiently transfected with the OVA OE pcDNA3 plasmid to construct target cells. OT-I CD8 + T cells were cocultured with either a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -Ctrl-mCherry cells or a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -KO-mCherry cells for 24 h ( n = 5 independent experiments per group). e Quantification of tumor-infiltrating CD4 + T cells and CD8 + T cells in subcutaneous KPC Scr/KD1 model ( n = 5 mice per group) and Panc02 Scr/KD1 model ( n = 9 mice per group). f Tumor growth curves from CD4 + or CD8 + T-cell deletion assay ( n = 5 mice per group). g Tumor growth evaluation in humanized NCG mice. Seven days post-PBMC injection, these immunoreconstituted mice were subcutaneously engrafted with Capan-1-Scr or Capan-1-KD1 cells. The level of immune humanization was confirmed by flow cytometric analysis of hCD45 + cells in peripheral blood on days 7 and 21. All mice were sacrificed 24 days after tumor inoculation for tumor weight measurement ( n = 7 mice per group). Statistical significance was determined via the unpaired t test (two-tailed) in ( c – g ). The data are presented as the means ± SDs. Source data are provided in the Source Data file.
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    a Hypothesis diagram depicting the crosstalk between immune inflammatory cells and PDAC cells. b Heatmap of transcriptome DEGs in the cellular response to IFN-γ process between KPC-shScramble (Scr) cells and KPC-shUbe2d3-1 (KD1) cells ( n = 3 samples per group). c Tumor growth curves following <t>subcutaneous</t> <t>inoculation</t> of KPC/Panc02 Scr/KD1 cells in syngeneic immunodeficient Rag1 −/− mice and subsequent implantation into immunocompetent C57BL/6 mice. Once tumor reached 1000 mm 3 in Rag1 −/− mice ( n = 4 mice per group), they were excised, and 1–2 mm 3 fragments were further inoculated subcutaneously into immunocompetent C57BL/6 mice ( n = 7 mice per group). d Experimental setup for in vitro CTL-mediated killing of KPC-WT/KO cells. KPC-WT cells were subjected to lentiviral transduction with the indicated sgCtrl <t>(Ctrl)-GFP,</t> sgCtrl (Ctrl)-mCherry, sgUbe2d3-1 (KO1)-mCherry, or sgUbe2d3-2 (KO2)-mCherry Cas9 vectors. These cells were further transiently transfected with the OVA OE pcDNA3 plasmid to construct target cells. OT-I CD8 + T cells were cocultured with either a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -Ctrl-mCherry cells or a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -KO-mCherry cells for 24 h ( n = 5 independent experiments per group). e Quantification of tumor-infiltrating CD4 + T cells and CD8 + T cells in subcutaneous KPC Scr/KD1 model ( n = 5 mice per group) and Panc02 Scr/KD1 model ( n = 9 mice per group). f Tumor growth curves from CD4 + or CD8 + T-cell deletion assay ( n = 5 mice per group). g Tumor growth evaluation in humanized NCG mice. Seven days post-PBMC injection, these immunoreconstituted mice were subcutaneously engrafted with Capan-1-Scr or Capan-1-KD1 cells. The level of immune humanization was confirmed by flow cytometric analysis of hCD45 + cells in peripheral blood on days 7 and 21. All mice were sacrificed 24 days after tumor inoculation for tumor weight measurement ( n = 7 mice per group). Statistical significance was determined via the unpaired t test (two-tailed) in ( c – g ). The data are presented as the means ± SDs. Source data are provided in the Source Data file.
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    a Hypothesis diagram depicting the crosstalk between immune inflammatory cells and PDAC cells. b Heatmap of transcriptome DEGs in the cellular response to IFN-γ process between KPC-shScramble (Scr) cells and KPC-shUbe2d3-1 (KD1) cells ( n = 3 samples per group). c Tumor growth curves following <t>subcutaneous</t> <t>inoculation</t> of KPC/Panc02 Scr/KD1 cells in syngeneic immunodeficient Rag1 −/− mice and subsequent implantation into immunocompetent C57BL/6 mice. Once tumor reached 1000 mm 3 in Rag1 −/− mice ( n = 4 mice per group), they were excised, and 1–2 mm 3 fragments were further inoculated subcutaneously into immunocompetent C57BL/6 mice ( n = 7 mice per group). d Experimental setup for in vitro CTL-mediated killing of KPC-WT/KO cells. KPC-WT cells were subjected to lentiviral transduction with the indicated sgCtrl <t>(Ctrl)-GFP,</t> sgCtrl (Ctrl)-mCherry, sgUbe2d3-1 (KO1)-mCherry, or sgUbe2d3-2 (KO2)-mCherry Cas9 vectors. These cells were further transiently transfected with the OVA OE pcDNA3 plasmid to construct target cells. OT-I CD8 + T cells were cocultured with either a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -Ctrl-mCherry cells or a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -KO-mCherry cells for 24 h ( n = 5 independent experiments per group). e Quantification of tumor-infiltrating CD4 + T cells and CD8 + T cells in subcutaneous KPC Scr/KD1 model ( n = 5 mice per group) and Panc02 Scr/KD1 model ( n = 9 mice per group). f Tumor growth curves from CD4 + or CD8 + T-cell deletion assay ( n = 5 mice per group). g Tumor growth evaluation in humanized NCG mice. Seven days post-PBMC injection, these immunoreconstituted mice were subcutaneously engrafted with Capan-1-Scr or Capan-1-KD1 cells. The level of immune humanization was confirmed by flow cytometric analysis of hCD45 + cells in peripheral blood on days 7 and 21. All mice were sacrificed 24 days after tumor inoculation for tumor weight measurement ( n = 7 mice per group). Statistical significance was determined via the unpaired t test (two-tailed) in ( c – g ). The data are presented as the means ± SDs. Source data are provided in the Source Data file.
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    a Hypothesis diagram depicting the crosstalk between immune inflammatory cells and PDAC cells. b Heatmap of transcriptome DEGs in the cellular response to IFN-γ process between KPC-shScramble (Scr) cells and KPC-shUbe2d3-1 (KD1) cells ( n = 3 samples per group). c Tumor growth curves following <t>subcutaneous</t> <t>inoculation</t> of KPC/Panc02 Scr/KD1 cells in syngeneic immunodeficient Rag1 −/− mice and subsequent implantation into immunocompetent C57BL/6 mice. Once tumor reached 1000 mm 3 in Rag1 −/− mice ( n = 4 mice per group), they were excised, and 1–2 mm 3 fragments were further inoculated subcutaneously into immunocompetent C57BL/6 mice ( n = 7 mice per group). d Experimental setup for in vitro CTL-mediated killing of KPC-WT/KO cells. KPC-WT cells were subjected to lentiviral transduction with the indicated sgCtrl <t>(Ctrl)-GFP,</t> sgCtrl (Ctrl)-mCherry, sgUbe2d3-1 (KO1)-mCherry, or sgUbe2d3-2 (KO2)-mCherry Cas9 vectors. These cells were further transiently transfected with the OVA OE pcDNA3 plasmid to construct target cells. OT-I CD8 + T cells were cocultured with either a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -Ctrl-mCherry cells or a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -KO-mCherry cells for 24 h ( n = 5 independent experiments per group). e Quantification of tumor-infiltrating CD4 + T cells and CD8 + T cells in subcutaneous KPC Scr/KD1 model ( n = 5 mice per group) and Panc02 Scr/KD1 model ( n = 9 mice per group). f Tumor growth curves from CD4 + or CD8 + T-cell deletion assay ( n = 5 mice per group). g Tumor growth evaluation in humanized NCG mice. Seven days post-PBMC injection, these immunoreconstituted mice were subcutaneously engrafted with Capan-1-Scr or Capan-1-KD1 cells. The level of immune humanization was confirmed by flow cytometric analysis of hCD45 + cells in peripheral blood on days 7 and 21. All mice were sacrificed 24 days after tumor inoculation for tumor weight measurement ( n = 7 mice per group). Statistical significance was determined via the unpaired t test (two-tailed) in ( c – g ). The data are presented as the means ± SDs. Source data are provided in the Source Data file.
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    a Hypothesis diagram depicting the crosstalk between immune inflammatory cells and PDAC cells. b Heatmap of transcriptome DEGs in the cellular response to IFN-γ process between KPC-shScramble (Scr) cells and KPC-shUbe2d3-1 (KD1) cells ( n = 3 samples per group). c Tumor growth curves following <t>subcutaneous</t> <t>inoculation</t> of KPC/Panc02 Scr/KD1 cells in syngeneic immunodeficient Rag1 −/− mice and subsequent implantation into immunocompetent C57BL/6 mice. Once tumor reached 1000 mm 3 in Rag1 −/− mice ( n = 4 mice per group), they were excised, and 1–2 mm 3 fragments were further inoculated subcutaneously into immunocompetent C57BL/6 mice ( n = 7 mice per group). d Experimental setup for in vitro CTL-mediated killing of KPC-WT/KO cells. KPC-WT cells were subjected to lentiviral transduction with the indicated sgCtrl <t>(Ctrl)-GFP,</t> sgCtrl (Ctrl)-mCherry, sgUbe2d3-1 (KO1)-mCherry, or sgUbe2d3-2 (KO2)-mCherry Cas9 vectors. These cells were further transiently transfected with the OVA OE pcDNA3 plasmid to construct target cells. OT-I CD8 + T cells were cocultured with either a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -Ctrl-mCherry cells or a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -KO-mCherry cells for 24 h ( n = 5 independent experiments per group). e Quantification of tumor-infiltrating CD4 + T cells and CD8 + T cells in subcutaneous KPC Scr/KD1 model ( n = 5 mice per group) and Panc02 Scr/KD1 model ( n = 9 mice per group). f Tumor growth curves from CD4 + or CD8 + T-cell deletion assay ( n = 5 mice per group). g Tumor growth evaluation in humanized NCG mice. Seven days post-PBMC injection, these immunoreconstituted mice were subcutaneously engrafted with Capan-1-Scr or Capan-1-KD1 cells. The level of immune humanization was confirmed by flow cytometric analysis of hCD45 + cells in peripheral blood on days 7 and 21. All mice were sacrificed 24 days after tumor inoculation for tumor weight measurement ( n = 7 mice per group). Statistical significance was determined via the unpaired t test (two-tailed) in ( c – g ). The data are presented as the means ± SDs. Source data are provided in the Source Data file.
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    Image Search Results


    a Hypothesis diagram depicting the crosstalk between immune inflammatory cells and PDAC cells. b Heatmap of transcriptome DEGs in the cellular response to IFN-γ process between KPC-shScramble (Scr) cells and KPC-shUbe2d3-1 (KD1) cells ( n = 3 samples per group). c Tumor growth curves following subcutaneous inoculation of KPC/Panc02 Scr/KD1 cells in syngeneic immunodeficient Rag1 −/− mice and subsequent implantation into immunocompetent C57BL/6 mice. Once tumor reached 1000 mm 3 in Rag1 −/− mice ( n = 4 mice per group), they were excised, and 1–2 mm 3 fragments were further inoculated subcutaneously into immunocompetent C57BL/6 mice ( n = 7 mice per group). d Experimental setup for in vitro CTL-mediated killing of KPC-WT/KO cells. KPC-WT cells were subjected to lentiviral transduction with the indicated sgCtrl (Ctrl)-GFP, sgCtrl (Ctrl)-mCherry, sgUbe2d3-1 (KO1)-mCherry, or sgUbe2d3-2 (KO2)-mCherry Cas9 vectors. These cells were further transiently transfected with the OVA OE pcDNA3 plasmid to construct target cells. OT-I CD8 + T cells were cocultured with either a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -Ctrl-mCherry cells or a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -KO-mCherry cells for 24 h ( n = 5 independent experiments per group). e Quantification of tumor-infiltrating CD4 + T cells and CD8 + T cells in subcutaneous KPC Scr/KD1 model ( n = 5 mice per group) and Panc02 Scr/KD1 model ( n = 9 mice per group). f Tumor growth curves from CD4 + or CD8 + T-cell deletion assay ( n = 5 mice per group). g Tumor growth evaluation in humanized NCG mice. Seven days post-PBMC injection, these immunoreconstituted mice were subcutaneously engrafted with Capan-1-Scr or Capan-1-KD1 cells. The level of immune humanization was confirmed by flow cytometric analysis of hCD45 + cells in peripheral blood on days 7 and 21. All mice were sacrificed 24 days after tumor inoculation for tumor weight measurement ( n = 7 mice per group). Statistical significance was determined via the unpaired t test (two-tailed) in ( c – g ). The data are presented as the means ± SDs. Source data are provided in the Source Data file.

    Journal: Nature Communications

    Article Title: IFN-γ-driven UBE2D3 upregulation impairs antigen presentation pathways and anti-tumor immunity in pancreatic cancer

    doi: 10.1038/s41467-025-65762-4

    Figure Lengend Snippet: a Hypothesis diagram depicting the crosstalk between immune inflammatory cells and PDAC cells. b Heatmap of transcriptome DEGs in the cellular response to IFN-γ process between KPC-shScramble (Scr) cells and KPC-shUbe2d3-1 (KD1) cells ( n = 3 samples per group). c Tumor growth curves following subcutaneous inoculation of KPC/Panc02 Scr/KD1 cells in syngeneic immunodeficient Rag1 −/− mice and subsequent implantation into immunocompetent C57BL/6 mice. Once tumor reached 1000 mm 3 in Rag1 −/− mice ( n = 4 mice per group), they were excised, and 1–2 mm 3 fragments were further inoculated subcutaneously into immunocompetent C57BL/6 mice ( n = 7 mice per group). d Experimental setup for in vitro CTL-mediated killing of KPC-WT/KO cells. KPC-WT cells were subjected to lentiviral transduction with the indicated sgCtrl (Ctrl)-GFP, sgCtrl (Ctrl)-mCherry, sgUbe2d3-1 (KO1)-mCherry, or sgUbe2d3-2 (KO2)-mCherry Cas9 vectors. These cells were further transiently transfected with the OVA OE pcDNA3 plasmid to construct target cells. OT-I CD8 + T cells were cocultured with either a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -Ctrl-mCherry cells or a mixture of KPC OVA -Ctrl-GFP cells and KPC OVA -KO-mCherry cells for 24 h ( n = 5 independent experiments per group). e Quantification of tumor-infiltrating CD4 + T cells and CD8 + T cells in subcutaneous KPC Scr/KD1 model ( n = 5 mice per group) and Panc02 Scr/KD1 model ( n = 9 mice per group). f Tumor growth curves from CD4 + or CD8 + T-cell deletion assay ( n = 5 mice per group). g Tumor growth evaluation in humanized NCG mice. Seven days post-PBMC injection, these immunoreconstituted mice were subcutaneously engrafted with Capan-1-Scr or Capan-1-KD1 cells. The level of immune humanization was confirmed by flow cytometric analysis of hCD45 + cells in peripheral blood on days 7 and 21. All mice were sacrificed 24 days after tumor inoculation for tumor weight measurement ( n = 7 mice per group). Statistical significance was determined via the unpaired t test (two-tailed) in ( c – g ). The data are presented as the means ± SDs. Source data are provided in the Source Data file.

    Article Snippet: The tumor burdens in the pancreas and liver were examined 21 d after tumor inoculation via IVIS detection of GFP signals (PerkinElmer, Lumina Series III).

    Techniques: In Vitro, Transduction, Transfection, Plasmid Preparation, Construct, DNA Deletion Assay, Injection, Two Tailed Test