Journal: EMBO Molecular Medicine
Article Title: TLR3 activation enhances abscopal effect of radiotherapy in HCC by promoting tumor ferroptosis
doi: 10.1038/s44321-024-00068-4
Figure Lengend Snippet: ( A ) ELISpot assay showing neoantigen specific-reactivity of splenic T cells against neoantigen peptide-pool with 7 mutant neoantigens identified in HCC Hepa1-6 cell line, and the results of three replicates were shown in statistical bar chart (right panel). The differences among the groups were analyzed by ANOVA. ( B ) The proportion of CD80 + CD86 + cells (gated on CD11c) in Tumor1 of different treated group as indicated was shown by representative flow cytometry scatter plots (left panel). The results of three replicates were shown in statistical scatter plots (right panel), and the differences among groups were analyzed by ANOVA. ( C ) The representative immunofluorescence image of CD45 + CD11c + CLEC9 + cDC1s in Tumor1 of different treated group (left panel), and the cell density of cDC1s (CD45 + CD11c + CLEC9 + cells) and IL12 + cDC1s of entire section field of the tumor tissue samples were shown in statistical scatter plots ( n = 3 slides from different mouse per group, right panel). Scale bars, 20 μm (400×). The differences between two groups were analyzed by t-test. ( D ) Flow cytometry showing the percentage of CD8 + central memory T cells in spleen from different treated group as indicated (left panel). The results of three replicates were shown in statistical scatter plots (right panel), and the differences among groups were analyzed by ANOVA. Data information: In ( A – D ), statistical data were presented as mean ± SEM (error bar) of three independent experiments. .
Article Snippet: Single-cell suspensions from tumor and monocytes derived from lymph nodes and spleen were blocked in 5%BSA (in PBS) or Fcblock for 20 min. For flow cytometry analysis of DC maturity, the monocytes derived from lymph nodes were stained in 0.5% BSA for 30 min with anti-mouse CD11c-PE mAb (1:100 dilution; eBioscience™, Waltham, USA; RRID: AB_465552), anti-mouse CD80-APC mAb (1:200 dilution; eBioscience™; RRID: AB_469417), anti-mouse CD86-PE/Cy7 mAb (1:100 dilution; eBioscience™; RRID: AB_2573372).
Techniques: Enzyme-linked Immunospot, Mutagenesis, Flow Cytometry, Immunofluorescence
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