GSM9025476: Control, D3, +/+; Mus musculus; RNA-Seq
Single cells from differentiated iHA-Etv2 embryoid bodies (EBs) at days 2, 3, and 4 (with and without doxycycline treatment) were isolated by FACS based on FLK1 and PDGFRα expression. Flow cytometry staining was performed using fluorophore-conjugated antibodies: FLK1-APC and PDGFRα-PE. Propidium iodide (PI) was used to exclude non-viable cells. A minimum of 10,000 viable cells were collected per condition for single-cell RNA sequencing. Single-cell RNA-seq was performed using the 10x Genomics Chromium Single Cell platform. Cell barcoding and cDNA library preparation were carried out following the manufacturer's protocol at the University of Minnesota Genomics Center (UMGC). Library quality was assessed using an Agilent Bioanalyzer High Sensitivity DNA chip. Sequencing was performed on an Illumina NovaSeq S4 platform with 2 × 150 bp reads, targeting approximately 50,000 reads per cell.