GSM5665210: Angptl4-knockout Day-1 post wounding; Mus musculus; RNA-Seq
Skin tissues were dissociated using 1mg/ml Type 3 collagenase and 10U/ml DNAseI for 1 hour in a 37°C water bath. The cells were strained through a 30 μm strainer and washed with PBS. Cells were blocked with 3% BSA containing Fc Blocker for 30 minutes, and then stained with CD45-APC (pan-hematopoietic cell marker) and propidium iodide (Live/Dead stain) for 1 hour on ice. After staining, cells were washed twice with PBS before sorting on the FACSAria Fusion cell sorter. Live immune cells (CD45+ PI-) cells were collected, washed twice with PBS, and counted on a hemocytometer before proceeding to single cell library preparation. Single cell droplets and cDNA libraries were prepared using the 10X Genomics Chromium Single Cell 3' V3 kit together with the Chromium Controller as per manufacturer's instructions.