Cd7 bv711

Cryopreserved immune cells were thawed and enriched for NK cells using the EasySep™ Human NK Cell Enrichment Kit (STEMCELL Technologies, Cambridge, MA). Enriched NK cells were stained with the following surface markers for flow cytometry: Live/Dead Blue, CD56 APC, CD3 BUV395, CD19 BV510 (BD, Franklin Lakes, NJ), CD18 APC-Cy7, CD7 BV711, CD16 BV605, and IFITM1 FITC. Cells were then stimulated with the following for 15 min: 500U/ml IL-2, 50ng/ml IL-12, 500ng/ml LPS, 500U/ml IFNα_a4, and 1ug/ml α-mouse IgG. After cytokine stimulation, cells were fixed with 4% paraformaldehyde for 10 min and then permeabilized with methanol for 30 min. Samples were then stained with the following intracellular antibodies overnight: pRPS6 BV421 (Biolegend, San Diego, CA). Samples were ran on a 5-laser Aurora (Cytek Biosciences, Fremont, CA).

Cryopreserved immune cells were thawed and enriched for NK cells using the EasySep™ Human NK Cell Enrichment Kit (STEMCELL Technologies, Cambridge, MA). Enriched NK cells were cultured with or without K562 cells at a 1:1 concentration for 4 hours with Golgi Stop added after 1 hour. Cells were then prepared for flow cytometry with the following markers: Live/Dead Blue (Invitrogen, Waltham, MA), CD56 APC, CD18 APC-Cy7, CD16 BV605 (Biolegend, San Diego, CA), CD3 BUV395, CD7 BV711, CD107 PE (BD, Franklin Lakes, NJ), and IFITM1 FITC (Biorbyt, St. Louis, MO).