Tripartite Separation of Glomerular Cell Types and Proteomes from Reporter-Free Mice.

ABSTRACT:

Background

The glomerulus comprises podocytes, mesangial cells, and endothelial cells, which jointly determine glomerular filtration. Understanding this intricate functional unit beyond the transcriptome requires bulk isolation of these cell types for biochemical investigations. We developed a globally applicable tripartite isolation method for murine mesangial and endothelial cells and podocytes (timMEP).

Methods

We separated glomerular cell types from wild-type or mT/mG mice via a novel FACS approach, and validated their purity. Cell type proteomes were compared between strains, ages, and sex. We applied timMEP to the podocyte-targeting, immunologic, THSD7A-associated, model of membranous nephropathy.

Results

timMEP enabled protein-biochemical analyses of pod

SUBMITTER: Hatje FA 

PROVIDER: S-EPMC8729851 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature