The experiment was performed using the same procedure as the 22-week heart unless noted here. The antibodies used for staining were PerCP/Cy5.5 CD235a (Biolegend #349110), PerCP/Cy5.5 CD45 (Biolegend #304028), FITC CD36 (Biolegend #336204), APC/Cy7 PECAM1 (Biolegend #303119). The gates were set up to sort cells with low DAPI, low CD45 (hematopoietic cells), low CD235A (erythroid cells), high PECAM1 (endothelial marker), and either low or high FITC. A total of 1824 PECAM1+ CD36-, PECAM1+ CD36+ and PECAM1+ cells from the 11-week heart and 1920 PECAM1+ CD36-, PECAM1+ CD36+ and PECAM1+ cells from the 14-week heart were sorted and processed for cDNA synthesis. A total of 1530 11-week and 1272 14-week cells that had sufficient cDNA concentration were barcoded and pooled for sequencing.
Synthesis of cDNA and library preparation for the fetal human heart cells was performed using the Smart-seq2 method as previously described (Picelli et al., 2014 (link); Su et al., 2018 (link)). Libraries from the fetal human heart cells were part of a pool of samples that was sequenced on four lanes of a Illumina NovaSeq 6000 S4 flow cell.
Synthesis of cDNA and library preparation for the fetal human heart cells was performed using the Smart-seq2 method as previously described (Picelli et al., 2014 (link); Su et al., 2018 (link)). Libraries from the fetal human heart cells were part of a pool of samples that was sequenced on four lanes of a Illumina NovaSeq 6000 S4 flow cell.
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