Anti cd8

The 29 NPC samples in our institutional cohort were divided into low and high expression groups based on the median value of IHC score of KIF18B. Eight samples were randomly selected from each group and conducted immunofluorescence staining. Briefly, the steps before incubation of secondary antibody were same as described in the above IHC protocol. The following primary antibodies were used: anti-FOXP3 (1:200, 98377S, Cell Signaling Technology), anti-CD8 (1:100, ZA-0508, OriGene), and anti-IBA-1 (1:200, 17198, Cell Signaling Technology). After incubation of primary antibodies, the slides were incubated with DyLight 488-labeled secondary antibody (1:500, ab96899, Abcam) in the dark at 37°C for 1h. Images were captured using immunofluorescence microscopy (Olympus, Tokyo, Japan). The quantification of cell numbers was carried out manually by counting the positive cells, while the immunofluorescence area was automatically identified using Image Pro Plus software (version 6.0.0.260).