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. 2008 Sep;19(9):3944–3955. doi: 10.1091/mbc.E08-04-0344

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© 2008 by The American Society for Cell Biology

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Figure 7. — Separated SN1 and SN2 form SDS-resistant complexes. (A and B) Cs (cerSN1) and sYs (citSN2) were expressed in HEK293T cells alone or together, with the indicated combinations of syntaxin and VAMP. Membranes were prepared as in Materials and Methods, and analyzed by SDS-PAGE followed by immunoblotting. In addition, FRET spectra were determined for some of these samples (see Figure 8). (A) Anti-GFP. (B) Anti-syntaxin. (C) anti-FLAG-VAMP. Complexes summarized in Table 1 are indicated by lower case letters: a and b, cerSN1 and syntaxin; c and d, cerSN1, syntaxin and VAMP; e, cerSN1, citSN2, syntaxin, and VAMP. Note that d is poorly resolved from b on these blots, but d contains VAMP (panel C, lane 2), whereas b does not (panel A, lane 2). (D) CerSN1 and mycSN2 were coexpressed with syntaxin and FLAG-VAMP in HEK293T cells. Membranes were analyzed by SDS-PAGE followed by immunoblotting. A complex containing all four proteins (designated f) was recognized by each of the indicated antibodies.