FL cells were thawed and allowed to recover at 37°C for an hour prior to staining.
Cells were blocked with TruStain FcX (BioLegend, 422301) and stained with TotalSeq A antibody mix containing 1ug of each TotalSeq A antibody per sample (BioLegend, see Table S1 for a list of antibodies). Anti-human CD235a-APC antibody (BD Biosciences, 551336) was added to the CD34 -sample. The CD34 + fraction was stained with anti-human CD34-APC antibody (BD Biosciences, 555824) and anti-human GPI-80-PE antibody (MBL International, D087-5). All samples were stained with calcein blue (Invitrogen, C34853) for live/dead exclusion and cell populations were sorted as shown in Figure S1A prior to loading onto the 10X Genomics platform. Chromium Single Cell 3ʹ Reagent Kit v3 with Feature Barcoding technology for Cell Surface Protein was used and the recommendations from the manufacturer were followed as specified in 10x
Genomics user guide document number CG000185, Rev B.
Cells were blocked with TruStain FcX (BioLegend, 422301) and stained with TotalSeq A antibody mix containing 1ug of each TotalSeq A antibody per sample (BioLegend, see Table S1 for a list of antibodies). Anti-human CD235a-APC antibody (BD Biosciences, 551336) was added to the CD34 -sample. The CD34 + fraction was stained with anti-human CD34-APC antibody (BD Biosciences, 555824) and anti-human GPI-80-PE antibody (MBL International, D087-5). All samples were stained with calcein blue (Invitrogen, C34853) for live/dead exclusion and cell populations were sorted as shown in Figure S1A prior to loading onto the 10X Genomics platform. Chromium Single Cell 3ʹ Reagent Kit v3 with Feature Barcoding technology for Cell Surface Protein was used and the recommendations from the manufacturer were followed as specified in 10x
Genomics user guide document number CG000185, Rev B.