Ficz 200nm

Sorted CD56^bright NK cells were cultured for 3 days on round-bottom 96-well plate, 10⁵ cells/well, in the presence of IL-2 100U/mL (NCI BRB Preclinical Repository) and either DMSO (ATCC), FICZ 200nM (Enzo Life Science) or CH-223191 3µM (Sigma-Aldrich). Then, culture supernatants were harvested, frozen at −80°C, and analyzed by 62-plex Luminex Immuno-Assay by the Human Immune Monitoring Center facility at Stanford University.

Our research was approved by the Institutional Review Board for human studies at Stanford University. Leukoreduction system (LRS) chambers were obtained from the Stanford Blood Center. NK cells were enriched using the RosetteSep™ NK Cell Enrichment Cocktail (STEMCELL Technologies) followed by centrifugation on Ficoll-Paque™ Premium (GE Healthcare). Then, CD56^bright and CD56^dim NK cells were sorted using the gating strategy showed on Supplemental Figure 1. NK cells were cultured on RPMI 1640 containing 10% FBS (Corning), 100U/mL Penicillin-Streptomycin, 55µM 2-Mercaptoethanol, 1x MEM Non-Essential Amino-Acids, 1mM Sodium Pyruvate, 10mM HEPES (Gibco). In order to stimulate AhR, before the culture, IL-2 100U/mL (NCI BRB Preclinical Repository) or IL-15 10ng/mL (Biolegend), and FICZ 200nM (Enzo Life Science) or CH-223191 3μM (Sigma-Aldrich) or DMSO (ATCC) as control were added to the media, as indicated.