GSM5770288: Pool-1, ADT, S4; Mus musculus; OTHER
LSK cells were enriched and processed as described above. Dead and apoptotic cells were removed using dead cell removal kit. Cells were blocked with TruStain FcX™ PLUS (anti-mouse CD16/32). Cell multiplexing and labeling was performed according to the manufacturer's instructions. Surface protein fixation was done by treating the cells with freshly prepared cell membrane crosslinker (DTSSP, 3,3'- dithiobis(sulfosuccinimidyl propionate). All samples were pooled in equal numbers into two groups (ambient air (~21% O2) and physioxia (3% O2)). Labeled cells were loaded to a 10X Genomics Chromium Next GEM chip G for reverse transcription and complementary DNA inside gel bead-in-emulsion. Full-length cDNA was amplified, and sequencing libraries were normalized for sequencing. The constructed library was sequenced on Illumina instrument. Library preparation was performed using New England Biolabs Single Cell/Low Input RNA Library Prep Kit for Illumina. Paired-end RNA-sequencing (RNA-seq) was performed on Illumina instrument by IU School of Medicine Medical Genomics core. Each group/condition consists of four matched biological replicates and ~30 million reads were obtained per sample.