Single-cell RNA-sequencing (scRNA-Seq) was performed on the 10x Genomics platform with Chromium Single Cell 3′ Chip Kit v2 (10x Genomics cat#1000009). Libraries were constructed using the Single Cell 3′ Library and Gel Bead Kit v2 (10x Genomics cat#120237) and Chromium i7 Multiplex Kit 10x Genomics cat#120262). Two single-cell libraries were pooled and sequenced per HiSeq 2500 125-base PET lane.
Conventional (bulk) RNA-sequencing was performed on unfractionated cell suspension or snap frozen whole tissue material. Total RNA was isolated with TRIzol reagent followed by purification over PureLink RNA Mini Kit columns (Invitrogen cat#12183018 A). RNA-seq was performed using a polyA-enriched strand-specific library construction protocol56 (link) and paired-end 75 bp sequencing on an Illumina HiSeq 2500 instrument.
Conventional (bulk) RNA-sequencing was performed on unfractionated cell suspension or snap frozen whole tissue material. Total RNA was isolated with TRIzol reagent followed by purification over PureLink RNA Mini Kit columns (Invitrogen cat#12183018 A). RNA-seq was performed using a polyA-enriched strand-specific library construction protocol56 (link) and paired-end 75 bp sequencing on an Illumina HiSeq 2500 instrument.
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