Figure 5. Functional proteins expressing on foxp3⁺ Treg in tumor-bearing mice spleen after VLP vaccination.

At 12^th day after tumor implantation and after one time VLP vaccination at day 3, mice were sacrificed for immune response analysis. About 10⁶ splenocytes were incubated with the appropriately diluted fluorescent-conjugated antibodies including CD4, foxp3, CTLA-4, CD25, CD44, PD-1, CD40L, and ICOS. FACSCalibur was used to acquire and FlowJo software was used to analyze the data. Cells were gated on CD4⁺foxp3⁺ population and histogram of different molecule expression in different experimental groups were presented in A). CTLA-4; B). CD25; C). CD44; D). PD-1; E). CD40L; F). ICOS. Shown were representative data from three experiments with at least three mice per group. (Statistically significances are shown, ** indicate p<0.01.).