Fig5. In situ localization of CD4+Foxp3+, CD8+Foxp3+, Foxp3+PD1 and Foxp3+PD-L1 inCC tissues(multiple fluorescence immunohistochemical staining).Representative images show tissue sections from CC with (A ) CD8+-opal650(magenta), Foxp3-opal570(pink), and DAPI(4,6-diamidino-2-phenylindole; blue) labeling, (B )CD4+-opal520(green), Foxp3-opal570(pink) and DAPI(blue) labeling, (C )Foxp3-opal570(pink), PD1-opal620(orange) and DAPI(blue) labeling, and (D )Foxp3-opal570(pink), PD-L1-opal540(yellow) and DAPI(blue) labeling. The graph show combined date for multiple fluorescence immunohistochemical staining analysis of counts of CD4+ /CD8+ T cell(E ), CD4+PD1/CD8+PD1(F ) and Foxp3+PD1/Foxp3+PD-L1(G ) expression on CC and healthy cervical tissues from the operation. Images(A ), (B ), (C ) and (D ) are from the same specimen and representative of CC patients. All images were captured using an Evos FL Auto 2 cell imaging system(Thermo Fisher Scientific) and merged using image J software. Data are reported as mean±SEM. *P<0.05;**P<0.01;***P<0.001;****P <0.0001(Student’s t test).