Fig6. In situ localization of CD4+, CD8+, Foxp3, PD1 and PD-L1 in CC tissues(multiple fluorescence immunohistochemical staining).Representative images show tissue sections from CC with CD4+(green), CD8+(magenta), Foxp3(pink), PD1(orange), PD-L1(yellow) and DAPI(blue) labeling. Looking at these images, it was found that the position presented by the Foxp3 staining almost coincides with that of the PD-L1, and the PD-L1 coincides with the location of tumor cell aggregation(A-C ). CD8+ T cell and PD1 were diffusely distributed, partially infiltrating tumor cell clusters and coincident with PD-L1 expression. Interestingly,the distribution of CD4+ T cell was almost around the tumor cell population. All images were captured using an Evos FL Auto 2 cell imaging system(Thermo Fisher Scientific) and merged using image J software.