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. Author manuscript; available in PMC: 2018 Mar 19.

Published in final edited form as: Cancer Immunol Res. 2017 Oct 15;5(11):1029–1045. doi: 10.1158/2326-6066.CIR-17-0175

Phenotyping of T cells. Circulating lymphocytes from a tumor-naïve C57BL/6 mice (left column) were compared with those from C57BL/6 mice treated and cleared of B16 primary tumors (right column) (n = 2 mice/group, representative of four independent experiments). Multiparametric flow cytometry for live CD4⁺ or CD8⁺ T cells (A), the fraction of CD4⁺ or CD8⁺ cells that are CD62L^hi or effector (CD62L^lo CD44^hi) phenotype (B and E), the fraction of CD62L^hi CD4⁺ or CD8⁺ cells expressing the inhibitory receptors (IR) PD-1 and TIM-3, the fraction of CD62L^lo CD44^hi effector cells expressing the IRs PD-1 and TIM-3 (D and G). To analyze quantitative flow cytometry data, one-way ANOVA testing was conducted with a Tukey posttest; P values reported from these analyses were corrected to account for multiple comparisons.

Phenotyping of T cells. Circulating lymphocytes from a tumor-naïve C57BL/6 mice (left column) were compared with those from C57BL/6 mice treated and cleared of B16 primary tumors (right column) (n = 2 mice/group, representative of four independent experiments). Multiparametric flow cytometry for live CD4⁺ or CD8⁺ T cells (A), the fraction of CD4⁺ or CD8⁺ cells that are CD62L^hi or effector (CD62L^lo CD44^hi) phenotype (B and E), the fraction of CD62L^hi CD4⁺ or CD8⁺ cells expressing the inhibitory receptors (IR) PD-1 and TIM-3, the fraction of CD62L^lo CD44^hi effector cells expressing the IRs PD-1 and TIM-3 (D and G). To analyze quantitative flow cytometry data, one-way ANOVA testing was conducted with a Tukey posttest; P values reported from these analyses were corrected to account for multiple comparisons.