FlowCytometryData_LN_B-NHL

LN-derived cells were thawed and stained for viability using a fixable viability dye e506 (Thermo Fisher Scientific, #65-0866-14) and for different surface markers depending on the experimental set-up. The following surface antibodies were used: anti-CD3-PerCP-Cy5.5, anti-CD4-PE-Dazzle, anti-CD8-APC-Cy7, anti-CD45RA-FITC, anti-CD25-BV421, anti-CD31-BV605, anti-CXCR5-BV711, anti-TIM3-BV711, anti-CD278-BV605, anti-PD1-PE-Cy7, anti-CD69-AF700, anti-CD244-BV421 (all BioLegend). For subsequent intracellular staining, cells were fixed and permeabilized with the intracellular fixation/permeabilization buffer set (Thermo Fisher Scientific, #88-8824-00) and stained with anti-Ki67-BV785, anti-FoxP3-AF647, anti-IKZF3-PE or adequate isotype controls (Thermo Fisher Scientific, BD Biosciences). Then, cells were analyzed using an LSR Fortessa (BD Biosciences) and FACSDiva (BD Biosciences, version 8). For analysis and gating of flow cytometry data FlowJo (v10.8.0) was used. Compensation was done prior measurement in FACSDiva. Corresponding compensation controls are provided for each measurement.