Annexin V-PI staining was performed using flow cytometric analysis as previously described50 (link). Briefly, 1 × 106 cells were cultured with increasing concentrations of selinexor (0–1000) for 24 h. Staining was performed using Apoptosis Detection Kit II (BD Biosciences, USA). Cells were harvested and washed twice with phosphate-buffered saline (PBS; Life technologies, USA). Cells were suspended in 1X binding buffer containing 5 ul of FITCI conjugated Annexin V and 5 ul of PI for 30 min in dark. The samples were analyzed using LSR-II flow cytometer (BD, San Jose, CA, USA).
Garg M., Kanojia D., Mayakonda A., Ganesan T.S., Sadhanandhan B., Suresh S., S. S., Nagare R.P., Said J.W., Doan N.B., Ding L.W., Baloglu E., Shacham S., Kauffman M, & Koeffler H.P. (2017). Selinexor (KPT-330) has antitumor activity against anaplastic thyroid carcinoma in vitro and in vivo and enhances sensitivity to doxorubicin. Scientific Reports, 7, 9749.
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