GSM9392655: CD107a- NK cells, nonresponsive; Homo sapiens; RNA-Seq
NK cells were isolated from fresh blood using a magnetic separation kit and rested overnight in complete RPMI supplemented with IL-2 (300 U/mL). Cells were cocultured with autologous EBV+ LCL for 4 hours at 37°C and 5% CO2. Cells were stained with Zombie Aqua, CD56-FITC (HCD56), CD3-PerCP-Cy5.5 (UCHT1) [Biolegend] and CD107a-efluor-660 (eBioH4A3) [eBioscience]. NK cells were sorted for live CD3−CD56+ and CD107a+/- on a BD FACSAria (BD Biosciences). Cells were washed in PBS and then pelleted for library preparation. Library was prepared according to the manufacturer's instructions using the 3' Chromium Next GEM Single Cell 3' GEM, Library & Gel Bead Kit v3.1 (10X Genomics).