Lack of a role of NHE4 in renal ammonia metabolism

Kidneys from a male and female mouse were used for single nucleus isolation and transcriptome analyses using the 10X Genomics platform. Overall design: Frozen kidney from mice were on the C57/Bl6 background strain and were homozygous for the floxed AR allele (AN Harris, RA Castro, HW Lee, JW Verlander, and ID Weiner. Role of the Renal Androgen Receptor in Sex Differences in Ammonia Metabolism. Am J Physiol Renal 321: F629-F44, 2021). The WT mice used for this paper were Cre-negative mice. Samples were homogenized with Nuclei lysis buffer plus RNAse inhibitors to generate single nucleus suspensions. Nuclei were loaded into Chromium Next GEM Chip G Single Cell Kit and libraries made with the Chromium Next GEM Single Cell 3' GEM, Library & Gel Bead Kit v3.1. Sequencing was performed with a targeted depth of 20K reads per nucleus on the NovaSeq 6000 with the UAB Genomics Core (Director Dr. Michael Crowley). Fastq files were mapped using mm10 genome, and fragments were counted with the 10X Genomics CellRanger pipeline (7.1.0).