Supp. Figure 3 data

After overnight recovery,<b> </b>purified T cells were stained with LIVE/DEAD Zombie fixable dead cell stain (0.2% in PBS) at RT in the dark for 15 minutes. T cells were washed twice in FACS buffer (PBS + 2% FCS) and stained in FACS buffer and 1% human Fc block (BD Pharmingen Human BD Fc Block) with mAb for the T cell surface markers CD4 BUV 395, CD8 BUV 805, CCR7 PE, CD45RA BUV496, CD62L PE-CF594, CD45RO PE-Cy7, and CD27 FITC (Supp. Figure 3B). Single colour compensation and fluorescence minus one (FMO) control samples were prepared for CCR7 and CD45RA. CD19+CD20+ target cell lines (Raji) were labelled with CD20+ and then stained with 0.2% LIVE/DEAD Zombie fixable dead cell stain, as above. T cells and target cells were washed twice after mAb staining in FACS buffer, resuspended at 2x10⁶ cells/ml in TCM, and co-incubated at a ratio of 1:1 +/- blinatumomab (10 ng/ml) for 1 hour at 37°C in a 5% CO₂ incubator. The samples were vortexed vigorously for 10 seconds and fixed by the addition of an equal volume of PBS containing 4% paraformaldehyde (Electron Microscopy Sciences, USA). Samples were acquired on a 5 laser Becton Dickinson LSR Fortessa (355, 405, 488, 561, 633 nm) conventional flow cytometer (BD Biosciences, USA).