Cell-surface protein
Cryopreserved peripheral blood mononuclear cells were profiled using scNTT-seq with cell-surface protein expression detection. The Biolegend TotalSeq-A oligo-conjugated antibody panel was used to profile cell-surface proteins, and a combination of two different barcoded nanobody-Tn5 fusion proteins used to profile H3K27me3 and H3K27ac genome-wide distribution. Single cells were captured using the 10x Genomics scATAC-seq kit. Protein expression (antibody-derived tag; ADT) and histone genomic DNA libraries were size-separated and amplified using separate PCRs. Reads were sequenced on a NextSeq 550 with the following sequencing format: i5: 38bp, i7: 8bp, read1: 60bp, read2: 60bp. Reads were demultiplexed into ADT and NTT-seq reads using the sample barcode, and different histone marks demultiplexed using the Tn5 adapter barcode sequence.