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. Author manuscript; available in PMC: 2017 Aug 1.
Published in final edited form as: Kidney Int. 2010 Jul 21;78(9):868–882. doi: 10.1038/ki.2010.212

Figure 1. Biochemical enrichment of slit diaphragm constituents.

Figure 1

Mouse glomeruli were isolated from kidney cortices, homogenized and fractionated to enrich for SD proteins. (a) The enrichment method is diagrammed schematically, indicating each fraction that was analyzed by immunoblotting (fractions 1–6). (b) An equal volume of extract was analyzed in lanes 2–6 to allow for a direct comparison between fractions. Replicate blots were probed with nephrin, podocin, podocalyxin, COX IV, PSMA2, WT1, flotillin 1, and transferrin receptor (TfR) antibodies (labeled to the right of each panel). As predicted, nephrin and podocin were located in the SD-enriched fraction, but podocalyxin, COX IV, PSMA2, and WT1 were absent from this fraction. Podocalyxin, COX IV, PSMA2, WT1, flotillin 1, and TfR receptor served as markers for apical membranes, mitochondria, cytoplasm, nuclei, lipid rafts, and nonraft domains, respectively.