Brief Method: kidneys from 6-week-old WT male mice were perfused via the left ventricle with perfusion buffer
(135 mM NaCl, 1 mM Na2HPO4, 1.2 mM Na2SO4, 1.2 mM MgSO4, 5 mM KCl, 2 mM CaCl2, 5.5 mM glucose, 5 mM HEPES, pH 7.4).
kidneys were then sliced and dissociated in perfusion buffer supplemented with collagenase B.
Microdissection were performed under the stereomicroscope equipped with on-stage cooling.
After washes in ice-cold PBS, the microdissected glomeruli were transferred to Trizol reagent for RNA extraction.
cDNAs were synthesized by SMARTer V4 kit (Clontech) and cDNA libraries were constructed for paired-end sequencing and sequenced on Illumina HiSeq3000 platform.
Reads were mapped to mouse Ensembl Genome by STAR and transcript abundances were calculated in the units of transcripts per million (TPM) using RSEM (https://github.com/deweylab/RSEM).
Please also visit
Mouse Renal Epithelial Cell Atals
.
Terminology:
G, Glomerulus. 5 glomerluli/sample from 3 mice.
Contact:
This website was created by Lihe Chen and Mark A. Knepper. Please contact:
knep@helix.nih.gov
.