gfp signals  (Revvity)

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

Journal: eLife

Article Title: Elevated DNA damage without signs of aging in the short-sleeping Mexican cavefish

doi: 10.7554/eLife.99191

Figure Lengend Snippet: ( A, B ) Immunostaining of cyclobutene pyrimidine dimer (CPD) shows a similar DNA damage level induced by UV in surface fish and Pachón cavefish embryonic fibroblasts. White circles indicate the nuclear area by DAPI staining. Orange indicates CPD. Scale bar, 40 μm. P -values were determined by two-way ANOVA: F =0.09703, p =0.7586. ns p =0.6404, **** p <0.0001. ( C, D ) Western blot of γH2AX indicates a diminished DNA damage response in Pachón cavefish embryonic fibroblasts compared to surface fish cells. ( E, F ) Flow cytometry images and quantification for host cell reactivation assays in surface fish and Pachón cavefish embryonic fibroblasts. Red line sets the green fluorescent protein (GFP) positive signal threshold. P -values were determined by unpaired t -test. ( G ) Representative GFP images for host cell reactivation assays in surface fish and Pachón cavefish embryonic fibroblasts. Scale bars, 500 μm. Figure 4—source data 1. Original gels for data presented in . Figure 4—source data 2. Original western blots for data presented in .

Article Snippet: Consistent with the γH2AX findings, the UV-damaged plasmid-transfected cavefish cells displayed a substantially lower GFP signal recovery (~22% relative to control) than the surface fish cells (~49% relative to control) ( ).

Techniques: Immunostaining, Staining, Western Blot, Flow Cytometry, Host-Cell Reactivation