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. 2016 Dec 10;8(3):4730–4746. doi: 10.18632/oncotarget.13972

Figure 6. Proportion of Treg and quantification of CD8+ T cells.

Figure 6

Figure 6

DLNs and tumor tissues were collected from mice treated with PBS, RdB, RdB/IL12, RdB/DCN, or RdB/IL12/DCN at 6 days after final viral treatment. (A) Population of Treg cells in DLNs from mice were analyzed by flow cytometry. Gating was for CD4+ T cells and analysis for CD25+ and Foxp3+ cells. Data points are mean ± SD of triplicate experiments with at least three mice per group. Similar results were obtained from at least two separate experiments. **P < 0.01, ***P < 0.001. (B) Cryosections of tumor tissues stained with anti-CD4 (green) and anti-Foxp3 (red) monoclonal antibodies. Arrows, T cells co-expressing CD4 and Foxp3 in tumors. Ratio of Foxp3+/CD4+ T cells was assessed by ImageJ software. The relative mean intensity of CD4- or Foxp3-positive cells were quantified from three independent fields within microscope images for each experimental group. Data are representative of three independent experiments. *P < 0.05, **P < 0.01, or ***P < 0.001. (C) Cryosections of tumor tissues were incubated with antibody against CD8 (green). Mean ± SD of CD8+ T cells was quantified by ImageJ software. The mean intensity of three different images was quantified for each sample (CD8+ T cells/field). Images are representative of three independent experiments. Original magnification: × 400. **P < 0.01.