Illumina HiSeq 4000 paired end sequencing; A single cell RNA-seq analysis of cardiac fibroblasts from pressure-overloaded mouse hearts
A single cell RNA-seq analysis of cardiac fibroblasts from pressure-overloaded mouse hearts
Eight-week-old wild-type C57BL/6 mice were subjected to pressure overload by means of transverse aortic constriction. Cardiac fibroblasts were isolated from heart ventricles 0, 1, and 3 days after the operation (cells from three mice were pooled in each sample). PI–, Fixable Viability Dye eFluor 450– CD45– CD31– MEFSK4+ single cells were sorted by FACS. processed for single-cell RNA sequencing analysis. Chromium 10X Genomics platform was used and approximately 15000 sorted single cell samples were loaded onto a Chromium controller (10x Genomics) for generation of gel bead-in-emulsions (GEM) using 10x Genomics Single Cell 3' Reagent Kit v3. Libraries were constructed using Single Cell 3' Reagent Kits v3 (10x Genomics)