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. Author manuscript; available in PMC: 2019 Aug 1.
Published in final edited form as: Kidney Int. 2018 May 31;94(2):326–345. doi: 10.1016/j.kint.2018.02.028

Figure 1. Isolation of glomerular endothelial cells from Flk1-H2B-EYFP mice.

Figure 1

(A) Kidney sections of Flk-1-H2B-EYFP mice were immunostained with antibodies against CD31 or WT-1 and counterstained with DAPI. Scale bar: 20μm. Rectangular area on the right panel is further magnified to visualize the colocalization of EYFP with CD31, but not with WT-1. (B) Single-cell suspension of isolated glomeruli from Flk1-H2B-EYFP mice was subjected to fluorescence-activated cell sorting (FACS) for EYFP+ cells. Typical FACS profile is shown. Approximately 15.7% EYFP+ GECs are obtained from dissociated glomerular cells. (C) Real-time PCR analysis of endothelial-specific or podocyte-specific genes show a robust enrichment of endothelial cell markers (Cdh5, Pecam1, and Icam1) in EYFP+ fraction, but were depleted of podocyte (Nphs1, Nphs2, and synaptopodin) and tubular cell (Pax8) markers as compared with EYFP- fraction.