SRX18026823 GSM6680846_r1 SRP404586 PRJNA894446 SRS15533979 GSM6680846 GSM6680846 RNA-Seq TRANSCRIPTOMIC SINGLE CELL cDNA Mouse aortas were cut into fine pieces and placed in 6.5 mL enzyme solution at 37°C for 1.5 hours (5 mL DMEM, 1 mL trypsin, collagenase type I 10 mg/mL, collagenase type IV 10 mg/mL , 50 µL elastin (2.5 mg/mL stock), collagenase type II 200 µL (stock: 1 mg/mL), Collagenase XI 200 µL (stock: 1 mg/mL), Hydrolase 200 µL (stock: 1 mg/mL), Liberase 100 µL (stock: 3.85 mg/mL, Roche), DNase I 10 µL (stock: 5U/µL, Sigma). This solution was then filtered using a 40 µm strainer (Falcon, NC), centrifuged (400xg for 5 min) and the cell pellet was suspended in 90 µL FEB buffer containing 0.5% BSA and 2 mM EDTA in PBS. 10 µL of CD31 Microbeads (MACS) was added to the suspension at 4°C for 15 min. Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics). Briefly, GCs were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the apropiated sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction Ilumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added. NextSeq 500 SRX18026824 GSM6680847_r1 SRP404586 PRJNA894446 SRS15533978 GSM6680847 GSM6680847 RNA-Seq TRANSCRIPTOMIC SINGLE CELL cDNA Mouse aortas were cut into fine pieces and placed in 6.5 mL enzyme solution at 37°C for 1.5 hours (5 mL DMEM, 1 mL trypsin, collagenase type I 10 mg/mL, collagenase type IV 10 mg/mL , 50 µL elastin (2.5 mg/mL stock), collagenase type II 200 µL (stock: 1 mg/mL), Collagenase XI 200 µL (stock: 1 mg/mL), Hydrolase 200 µL (stock: 1 mg/mL), Liberase 100 µL (stock: 3.85 mg/mL, Roche), DNase I 10 µL (stock: 5U/µL, Sigma). This solution was then filtered using a 40 µm strainer (Falcon, NC), centrifuged (400xg for 5 min) and the cell pellet was suspended in 90 µL FEB buffer containing 0.5% BSA and 2 mM EDTA in PBS. 10 µL of CD31 Microbeads (MACS) was added to the suspension at 4°C for 15 min. Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics). Briefly, GCs were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the apropiated sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction Ilumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added. NextSeq 500