scRNAseq of PBMCs of adolescents with Asthma
Cells were processed in batches of 16. For each batch, PBMCs were thawed and plated in a 96-well plate. Cells were incubated in starvation media overnight at 37C and 5% CO2. The following morning cells for each individuals were treated with either LPS and dexamethasone, LPS and vehicle control alone, PHA and  dexamethasone, PHA and vehicle control alone, or vehicle control alone. After six hours, cells were pooled across individuals for a total of five treatment-specific pools. Cell concentration was adjusted and cells with each treatment condition were loaded onto a separate channel of 10x Genomics Chromium instrument according to the manufacturer's protocol, with batches 4, 5 and 6 loaded on two separate Chromium Chips for a total of 2 wells per treatment pool. Batches 1-4 and one chip of batch 5 were processed using v2 chemistry, and the other chip of batch 5 and batch 6 were processed using v3 chemistry. Library preparation was done according to the manufacturer's protocol. Sequencing of the single-cell libraries was performed on an Illumina Nextseq 500 instrument.