Figure 7.

B16-F10 cells exposed to chromomycin A5 induce activation of CD4⁺ and CD8⁺ T lymphocytes and generate cell death in viable B16-F10 cells. Cells pre-incubated for 24 h with 0.1 μM CA₅ (CA5) and 0.6 μM doxorubicin (Dox), or sterile saline (C-) were injected subcutaneously in the right axilla of mice and their splenocytes were evaluated 7 days post-vaccination. (A) Representative contour plot graphs of CD69 surface marker on CD8 T lymphocytes. (B) Column graphs showing the percentage of CD8⁺CD69⁺ T lymphocytes. (C) Representative contour plot graphs of the CD69 surface marker on CD4 T lymphocytes. (D) Column graphs showing the percentage of CD4⁺CD69⁺ T lymphocytes. (E) Representative contour plot graphs of CD44 surface marker on CD4 T lymphocytes showing CD44^high and CD44^low populations and (F) illustration identifying CD44^high and CD44^low populations. (G) Column graphs showing the percentage of CD4⁺CD44^high T lymphocytes, CD4⁺CD44^low T and CD4⁺CD25⁺ T lymphocytes and (H) column graphs showing the percentage of CD8⁺CD25⁺ T lymphocytes. (I) Column graphs showing cell death of B16-F10 cells after 5h incubation with splenocytes ratios of effector/target (E/T). The splenocytes were previously restimulated with sterile saline (C-), CA5 and Dox for 72h and 120h before the cytotoxic assay. Differences between groups are expressed as p values indicated above the compared groups. N= 5 animals per group.