MOESM3 of Activation of natural killer cells by rituximab in granulomatosis with polyangiitis

Additional file 3: Figure S3. Gating strategy for measurement of in vivo NK cell activation. The gating has been performed in a standardized way, and a typical GPA patient is shown. a First, live cells were roughly gated based on forward and sideward scatter (FSC, SSC). Second, Zombie Aqua™ viability dye positive cells were determined as “dead” and remaining cells as “live”. As shown on the bottom, peripheral blood lymphocytes (PBL) were mostly in the live gate, and now re-gated in a conservative, “tight” fashion to exclude monocytes and, as good as possible, potentially apoptotic cells which would be located on the upper left part of the main population. b Among PBL, T cells were determined as CD3 + CD19-, B cells as CD3-CD19+ and NK cells as CD3-CD19-CD56+ cells. FMO (“fluorescence minus one”) controls were conducted in all experiments.