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. 1999 Jul 6;96(14):7962–7967. doi: 10.1073/pnas.96.14.7962

Figure 3.

Figure 3

Immunoelectron microscopic localization of nephrin in human renal glomeruli. Indirect postembedding staining for nephrin by using affinity purified IgG against extracellular region of recombinant human nephrin and 10 nm gold-coupled secondary antibody. (A) Paraformaldehyde (PF)-fixed kidney embedded in Lowicryl. Note gold label (arrowheads) between foot processes of podocytes (P). The label is located in the central area of the slit, between the GBM and the faintly visible slit diaphragm (arrows). Endothelium (E) is unlabeled. Bar = 200 nm. (B) Several gold particles in a row (box) can be seen between tangentially sectioned podocyte (P) foot processes. Gold particles in two other podocyte–podocyte interspaces are shown with arrowheads. Apparent intracellular staining of a podocyte (arrows) above GBM could be caused by grazing section of slit. Sample treated as in A. Bar = 500 nm. (C) Blow-up of B. The row of nine gold particles lies in tangentially cut, ca. 40-nm-wide slit between two podocytes (P). GBM is marked with asterisk. Bar = 50 nm. (D) Gold particle between slit diaphragm (arrow) joining two podocytes (P) and GBM (asterisk) in cross section. Sample fixed in 3.5% PF with 0.01% glutaraldehyde and embedded in LR White resin. Bar = 50 nm.