Experimental, Rep 2
After 7 days of co-incubation with cancer spheroids, NK cells were isolated with an NK cell selection kit (EasySep CD56+ selection kit; StemCell technologies) and analyzed by flow cytometry (Northern Lights 2000 Full Spectrum, Cytek) to determine NK cell purity. Total RNA was extracted from the NK cells (Direct-zol Microprep kit, Zymo research). RNA quality (RIN value) was determined by TapeStation and used for polyA selection, library preparation, and RNA sequencing (Genewiz, Inc, South Plainfield, NJ).