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. 2001 Oct;12(10):3031–3045. doi: 10.1091/mbc.12.10.3031

Figure 4.

Figure 4

IG EM localization of reggie-1 and -2, Thy-1, fyn, and caveolin-1. Orientation of sections is indicated in the insets. Bars, 0.1 μm. (a) Astrocyte, double labeling of sections with anti-reggie-2 pABs and pA-Au10, followed by anti-reggie-1 mAB and goat antimouse F(ab)2-Au5. The area shown is a “grazing” section along the plasma membrane as indicated in the inset. Double labeling occurs in small clusters at the cell periphery (arrowheads). (b) PC12 cell (after cross-linking of Thy-1 in vivo) double labeling on sections with anti-Thy-1 mAB and goat antimouse-Au10 followed by anti-reggie-2 pAB and pA-Au5. Double labeling occurs along apposed cell membranes (arrowheads) of two neighboring cells. S, support. (c) PC12 cell (after cross-linking of Thy-1 in vivo) double labeling on sections with anti-Thy-1 mAB and goat antimouse F(ab)2-Au5, followed by anti-reggie-2 pAB and pA-Au10. Note double labeling of ∼0.1-μm large microdomains on a grazing section along the plasma membrane. (d) DRG neurites [exposed to anti-fyn pAB and pA-Au10, followed by anti-reggie-1 mAB and goat antimouse F(ab)2-Au5]. Note colocalization (arrowheads) on a neurite. S, support. (e–g) Astrocyte, horizontal (grazing) section (e), and cross section (f) through caveolae and regions devoid of caveolae (g). Micrographs (e–g) are from the same section. Sections were subjected to double immunogold labeling with anti-caveolin-1 pAB and pA-Au10 and to anti-reggie-1 mAB and goat antimouse F(ab)2-Au5. Au10 gold grains label caveolae (e and f, arrowheads) where Au5 gold grains are not found. (g) Au5 gold grains indicating the presence of reggie-1 are detected in regions of the plasma membrane clearly distinct from caveolae and anti caveolin-1 (Au10) label.