Single cell RNASeq of whole mouse kidneys from Uromodulin/Tamm horsfall protein knock-out mice. Single cell RNASeq of whole mouse kidneys from Uromodulin/Tamm horsfall protein knock-out mice

To elucidate changes in cellular populations in the ischemic acute kidney injury, we performed single cell RNAseq on mouse kidneys following ischemic reperfusion injury and a sham surgery. Mice were subjected to bilateral clamping of the renal pedicule for 30 minutes. After 6-7 hours of reperfusion the kidneys were removed and processed by a MACs cell isolation protocol and dead cell removal (PMID:33448928) , sorted on a 10X Chromium platform version 3 and libraries generated. Sequencing was performed on an Illumina NovaSeq 6000. Overall design: Uromodulin/Tamm horsfall protein knock-out 129ev mice were subjected to a Sham surgery or bilateral renal clamping for 30 minutes.

为阐明缺血性急性肾损伤中细胞群体的变化特征，我们对缺血再灌注损伤组与假手术组的小鼠肾脏开展了单细胞RNA测序（single cell RNAseq）实验。实验小鼠接受双侧肾蒂钳夹30分钟处理，再灌注6-7小时后摘取肾脏，采用MACs细胞分离方案及死细胞清除流程（参照文献PMID:33448928）进行样本制备，随后通过10X Chromium V3平台进行细胞分选并构建测序文库。测序工作在Illumina NovaSeq 6000平台上完成。实验整体设计：选用尿调节素/塔姆-霍斯福尔蛋白（Tamm-Horsfall Protein）敲除的129ev系小鼠，分别接受假手术或双侧肾蒂钳夹30分钟处理。