Proximal tubular cells with different DNA contents single-cell sequencing

To explore the heterogeneity and functional dynamics of proximal tubular cells (PTCs) with different DNA contents, we isolated PTCs (2N, 4N, >4N) from male C57BL/6J mouse kidneys at day 3 and day 14 post-ischemia-reperfusion injury, and generated a single-cell transcriptome atlas.The renal cortices of mice were minced and digested, then the PTCs with different DNA contents were isolated to perform scRNA-seq by fluorescence-activated cell sorting (FACS) based on lotus tetragonolobus lectin (LTL)-positive and DNA content indicated by Hoechst 33342. Library construction was performed according to the instructions of 10x Genomics, and sequencing was conducted using the Illumina HiSeq 4000 system.