Cisplatin- and oxaliplatin-treated cells on slide chamber culture plates were washed and fixed with ice-cold methanol for 10 min, and then permeabilized with 0.1% Triton X-100 for 15 min. After blocking with 0.1% tween, cells were stained with anti-CRT Alexa Fluor® 488 (Abcam, Cambridge, UK) overnight. Cells were then counter-stained with DAPI and visualized under a fluorescent microscope.
For flow cytometer analysis, cisplatin- and oxaliplatin-treated cells were washed and stained with anti-CRT Alexa Fluor® 488 (Abcam, Cambridge, UK) for 1 h. The expression of ecto-CRT was analyzed using the CytoFLEX cell sorter (Beckman Coulter, Brea, CA, USA).
For flow cytometer analysis, cisplatin- and oxaliplatin-treated cells were washed and stained with anti-CRT Alexa Fluor® 488 (Abcam, Cambridge, UK) for 1 h. The expression of ecto-CRT was analyzed using the CytoFLEX cell sorter (Beckman Coulter, Brea, CA, USA).