For assessment of isolation purities, flow cytometry was conducted using the Cytoflex system (Beckman Coulter). For RNA-seq library validation experiments and benchtop comparisons, PBMCs were sorted using the MoFlo Astrios (Beckman Coulter). Lysate pools were generated by sorting 5000 cells into 20 μl of TCL buffer (Qiagen) and 1 μl proteinase K (20 mg/ml; Qiagen) and stored at −80°C to maintain RNA integrity. The following panel was used for both purity assessment and sorting: DAPI, CD45 BV605, CD3 AF700, CD4 FITC, CD8 PE, CD14 APC, CD19 PE-Cy7, and CD56 BV650 (all IgG1κ, BioLegend). Flow cytometry data were analyzed using FlowJo v10.1.

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