Annexin-V/PI staining assay (BD Biosciences, San Jose, CA) was performed as previously described32 (link). HN3 cells were plated at 1 × 106 cells/well in 6-well plates and were treated with TRAIL and/or VPA. After 24 h of cell seeding, the apoptotic cells were stained with Annexin-V-FITC and PI solution, and the stained cells were analyzed via flow cytometry (BD Biosciences).
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