Subunit composition of mammalian transient receptor potential channels in living cells -- Data Supplement - Supporting Figure -- Proceedings of the National Academy of Sciences

Supporting information for Hofmann et al. (May 14, 2002) Proc. Natl. Acad. Sci. USA, 10.1073/pnas.102596199.

Fig. 5.
Colocalization of fluorescent protein-tagged transient receptor potential channel (TRPC) constructs with a membrane marker. Live HEK 293 cells expressing (a) hTRPC6^wt-GFP, (b) hTRPC1-YFP, and (c) hTRPC6^D131-GFP were stained for 10 min with a DiI membrane-labeling solution (1% vol/vol; Vybrant, Molecular Probes) in Hepes-buffered saline (HBS; 140 mM NaCl/6 mM KCl/1.25 mM MgCl₂/1.25 mM CaCl₂/10 mM Hepes/5 mM glucose/0.1 % BSA, pH 7.4), washed twice in HBS, and subjected to confocal laser-scanning fluorescent microscopy with a Leica TCS-SP2 confocal microscope. Green and yellow fluorescent proteins (GFP/YFP) were excited at 488 nm, DiI at 543 nm. GFP/YFP emission was recorded between 500 and 530 nm, DiI emission above 560 nm. For each combination, the GFP/YFP fluorescence (Left), the DiI fluorescence (Center), and an overlay image of both (Right) are depicted. The localization of the plasma membrane (pm), the nuclear envelope (ne), and overlays (ovl) are indicated by arrows.