GSM6406546: wk10_control_Envpos; Macaca mulatta; RNA-Seq
Frozen FNA or PBMC samples were thawed and recovered in 50 % (v/v) FBS in RPMI. Recovered live cells were enumerated and stained with the appropriate staining panel. MD39 and MD39-base KO baits were prepared by mixing biotinylated MD39 with fluorophore-conjugated streptavidin (SA) in small increments at RT in an appropriate volume of 1X PBS over the course of 45 min. MD39:SA were added to the cells for 20 minutes, after which the antibody master mix was added for another 30 minutes at 4 ºC. Where KO baits were used, KO baits were added to the cells first for 20 minutes, then WT MD39:SA baits and were added for another 20 min, followed by the addition of the remainder of the staining panel for an additional 30 minutes at 4ºC, similar to previously described12. Fully supplemented RPMI (R10; 10 % (v/v) FBS, 1x pen/strep, 1x Glutamax) was used as FACS buffer. For sorting, anti-human hashtag antibodies (Biolegend) were individually added to each sample at a concentration of 2.5 µg/5 million cells at the time of addition of the master mix. Library construction was performed according to the manufacter's instructions (Chromium Next GEM Single Cell V(D)J Reagent Kit v1.1, 10x Genomics for  LN FNA samples and Chromium Next GEM Single Cell 5' Reagent Kit v2 (Dual Index), 10x Genomics for PBMC samples ).