GSM8580009: WT_Nfkb1-KO_BMDC_Day3, GEX; Mus musculus; OTHER
On day 3 or day 5, three wells of cells were harvested, pooled before staining with surface antibodies for CD3/CD19/CD49b/Ter119-FITC, Gr-1-PECy7, CD45.1-BV711, CD45.2 APCCy7 and DAPI. Live Dump-(CD3/CD19/CD49b/TER119/Gr-1-) CD45.1+CD45.2- (WT) cells and live Dump-CD45.1-CD45.2+ (Nfkb1-/-) cells were sorted by BD FACS Aria II (UCLA JCCC Flow Core). Sorted cells were stained with unique TotalSeq™-A anti-mouse Hashtag Antibodies (TotalSeq™-A #1,2,3,5 were used, BioLegend, San Diego, CA) for multiplexing. On each day, stained CD45.1 WT cells and CD45.2 Nfkb1-/- cells were washed once, counted, mixed at 1:1 ratio and resuspended in PBS with 10% fetal bovine serum while the concentration was adjusted to 1200 cells/μL. The cells were then submitted to UCLA Technology Center for Genomics & Bioinformatics for 10X Genomics (Pleasaton, CA) Chromium captures. Each independent sample was loaded into a separate GEM wells and TotalSeq™-A Antibodies and Cell Hashing with 10x Single Cell 3' Reagent Kit v3 protocol was applied.