For immunocytochemistry, cells were fixed in 4% paraformaldehyde at room temperature for 15 min, permeabilized in 5% Triton X-100 for 5 min and then stained using primary antibodies. The secondary antibodies used were anti-mouse Alexa Fluor 488 or 594 dye conjugate and/or anti-rabbit Alexa Fluor 488 or 594 dye conjugate (Life Technologies). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue; Life Technologies). After mounting, the cells were visualized using a multiphoton confocal laser-scanning microscope (Carl Zeiss, Thornwood, NY, USA).
Li C.W., Lim S.O., Xia W., Lee H.H., Chan L.C., Kuo C.W., Khoo K.H., Chang S.S., Cha J.H., Kim T., Hsu J.L., Wu Y., Hsu J.M., Yamaguchi H., Ding Q., Wang Y., Yao J., Lee C.C., Wu H.J., Sahin A.A., Allison J.P., Yu D., Hortobagyi G.N, & Hung M.C. (2016). Glycosylation and stabilization of programmed death ligand-1 suppresses T-cell activity. Nature Communications, 7, 12632.
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