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. Author manuscript; available in PMC: 2010 Dec 30.

Published in final edited form as: Vaccine. 2009 Dec 30;27(Suppl 6):G52–G59. doi: 10.1016/j.vaccine.2009.09.106

Figure 2a. Relatively increased numbers of Tregs in periphery of TRAMP mice with increasing age. Splenocytes isolated from groups of young (≤ 8 weeks old, n = 3), middle-aged (16–20 weeks old, n = 4) and old (≥ 24 weeks old, n = 3) TRAMP mice were washed and stained with anti-mouse CD3-FITC and anti-mouse CD4-PE/Cy7. Cells were fixed and permeablised overnight, then stained with anti-mouse FOXP3-PE and analysed by flow cytometry. Events were collected gated on live, CD3⁺ cells and then further gated on the CD4⁺FOXP3⁻ fraction and the CD4⁺FOXP3⁺ fraction. The * symbol indicates p < 0.05.

Figure 2b. Increased infiltration of CD4⁺ T cells and Treg into the prostate tumor with increasing age. Tumor infiltrating lymphocytes isolated from groups of young ( 8 weeks old, n = 3), middle-aged (16–20 weeks old, n = 4) and old ( 24 weeks old, n = 3) TRAMP mice were washed and stained with anti-mouse CD3-FITC, anti-mouse CD8-PE/Cy5 and anti-mouse CD4-PE/Cy7. Cells were fixed and permeablised overnight, then stained with anti-mouse FOXP3-PE and analysed by flow cytometry. Events were collected gated on live, CD3⁺CD8⁻ cells and then further gated on the total CD4⁺FOXP3⁻ fraction and the CD4⁺FOXP3⁺ fraction. The‡ symbol indicates p < 0.001.

Figure 2a. Relatively increased numbers of Tregs in periphery of TRAMP mice with increasing age. Splenocytes isolated from groups of young (≤ 8 weeks old, n = 3), middle-aged (16–20 weeks old, n = 4) and old (≥ 24 weeks old, n = 3) TRAMP mice were washed and stained with anti-mouse CD3-FITC and anti-mouse CD4-PE/Cy7. Cells were fixed and permeablised overnight, then stained with anti-mouse FOXP3-PE and analysed by flow cytometry. Events were collected gated on live, CD3⁺ cells and then further gated on the CD4⁺FOXP3⁻ fraction and the CD4⁺FOXP3⁺ fraction. The * symbol indicates p < 0.05.

Figure 2b. Increased infiltration of CD4⁺ T cells and Treg into the prostate tumor with increasing age. Tumor infiltrating lymphocytes isolated from groups of young ( 8 weeks old, n = 3), middle-aged (16–20 weeks old, n = 4) and old ( 24 weeks old, n = 3) TRAMP mice were washed and stained with anti-mouse CD3-FITC, anti-mouse CD8-PE/Cy5 and anti-mouse CD4-PE/Cy7. Cells were fixed and permeablised overnight, then stained with anti-mouse FOXP3-PE and analysed by flow cytometry. Events were collected gated on live, CD3⁺CD8⁻ cells and then further gated on the total CD4⁺FOXP3⁻ fraction and the CD4⁺FOXP3⁺ fraction. The‡ symbol indicates p < 0.001.